Compositions and methods for inhibiting TGF-beta

ABSTRACT

The present invention provides compounds of the formula:  
                 
 
and pharmaceutically acceptable isomers, salts, hydrates, solvates, and prodrug derivatives thereof, wherein R 1 , R 6 , R 7 , R 8 , Pg, and n are those defined herein. The present invention also provides pharmaceutical compositions comprising the same and methods for using the same. In particular, compounds of Formula I are useful in modulating TGF-β activity.

CROSSED-REFERENCE TO RELATED APPLICATION

This application claims the benefit of U.S. Provisional PatentApplication No. 60/479,488, filed Jun. 17, 2003, the content of which isincorporated herein by reference.

FIELD OF THE INVENTION

This invention relates to compounds that are useful in modulating thetransforming growth factor (TGF)-β signaling pathway.

BACKGROUND OF THE INVENTION

TGF-β belongs to a large super-family of multifunctional polypeptidefactors. The TGF-β family includes three genes, TGF-β1, TGF-β2 andTGF-β3, which are pleiotropic modulators of cell growth anddifferentiation, embryonic and bone development, extracellular matrixformation, hematopoiesis, immune and inflammatory responses. Forexample, TGF-β1 inhibits the growth of many cell types, includingepithelial cells, but stimulates the proliferation of various types ofmesenchymal cells.

The TGF-β genes have high homology with one another. In mammals, theTGF-β super-family includes various TGF-β genes, as well as theembryonic morphogenes, such as the family of the activins, inhibins,“Mullerian Inhibiting Substance”, and bone morphogenic protein (BMP).See Roberts and Spom, “The Transforming Growth Factor-βs in PeptideGrowth Factors and Their Receptors. I.” Handbook of ExperimentalPharmacology, vol. 95/I, Springer-Verlag, Berlin, 1990, pp 419-472. Eachmember of the TGF-β family exerts a wide range of biological effects ona large variety of cell types, e.g., they regulate cell growth,morphogenesis, differentiation, matrix production and apoptosis. Lagnaet al., Nature, 1996, 383, 832-836. TGF-β acts as a growth inhibitor formany cell types and is believed to play a central role in the regulationof embryonic development, tissue regeneration, immuno-regulation, aswell as in fibrosis and carcinogenesis.

In addition, TGF-β induces the synthesis of extracellular matrix (ECM)proteins, modulates the expression of matrix proteinases and proteinaseinhibitors and changes the expression of integrins. ECM is a dynamicsuperstructure of self aggregating macromolecules including fibronectin,collagen and proteoglycan. It is believed that ECM is the chiefpathologic feature of fibrotic diseases. ECM disorder has also beenproposed to play a central role in pathogenesis disorders such ashypertensive vascular disease and diabetic renal disease. Sato et al.,Am. J. Hypertens., 1995, 8, 160-166 (1995); Schulick et al., Proc. Natl.Acad. Sci., 1988, 95, 6983-6988. Moreover, TGF-β is expressed in largeamounts in many tumors. Derynck, Trends Biochem. Sci., 1994, 19,548-553. This strong occurrence in neoplastic tissues could indicatethat TGF-β is strategic growth/morphogenesis factor which influences themalignant properties associated with the various stages of themetastatic cascade. TGF-β inhibits the growth of normal epithelial andrelatively differentiated carcinoma cells, whereas undifferentiatedtumor cells which lack many epithelial properties are generallyresistant to growth inhibition by TGF-β. Hoosein et al., Exp. Cell.Res., 1989, 181, 442-453; Murthy et al., Int'l. J Cancer, 1989, 44,110-115. Furthermore, TGF-β1 is believed to potentiate the invasive andmetastatic potential of a breast adenoma cell line (Welch et al., Proc.Natl. Acad. Sci., 1990, 87, 7678-7682), which indicates a role of TGF-β1in tumor progression.

The cellular effects of TGF-β are exerted by ligand-inducedhetero-oligomerization of two distantly related type I and type IIserine/threonine kinase receptors, TGF-βR-I and TGF-βR-II, respectively.Lin et al., Trends Cell Biol., 1993, 11, 972-978; Massague et al.,Cancer Surv., 1996, 27, 41-64; Dijke et al., Curr. Opin. Cell Biol.,1996, 8, 139-145. The two receptors, both of which are required forsignaling, act in sequence: TGF βR-I is a substrate for theconstitutively active TGF-βR-II kinase. Wrana et al., Nature, 1994, 370,341-347; Wieser et al., EMBO J., 1995, 14, 2199-2208. Upon TGF-β1binding, the type II receptor phosphorylates threonine residues in GSdomain of ligand occupied type 1 receptor or activin like kinase (ALK5),which results in activation of type I receptors. The TGF-β1 type Ireceptor in turn phosphorylates Smad2 and Smad3 proteins whichtranslocate to the nucleus and mediate intracellular signaling. Theinhibition of ALK5 phosphorylation of Smad3 reduces TGF-β1 inducedextracellular matrix production. Krettzchmar et al., Genes Dev., 1997,11, 984-995; Wu et al., Mol. Cell. Biol., 1997, 17,2521-2528.

TGF-β is also a powerful and essential immune regulator in the vascularsystem capable of modulating inflammatory events in both leuko andvascular endothelial cells. Shull et al., Nature, 1992, 359, 693-699. Itis also involved in the pathogenesis of chronic vascular diseases suchas atherosclerosis and hypertension. Grainger & Metcalfe et al., Bio.Rev. Cambridge Phil. Soc., 1995, 70, 571-596; Metcalfe et al., J. HumanHypertens., 1995, 9, 679.

Genetic studies of TGF-β-like signaling pathways in Drosophila andCaenorhabditis elegans have led to the identification of mothers againstdpp (Mad) and sma genes, respectively. Sekelsky et al., Genetics, 1995,139, 1347-1358; Savage et al., Proc. Natl. Acad. Sci. USA, 1996, 93,790-794. The products of these related genes perform essential functionsdownstream of TGF-β like ligands acting via serine/threonine kinasereceptors in these organisms. Wiersdorf et al., Development, 1996, 122,2153-2163; Newfeld et al., Development, 1996, 122, 2099-2108; Hoodlesset al., Cell, 1996, 85, 489-500.

Vertebrate homologs of Mad and sma have been termed Smads or MADR genes.Derynck et al., Cell, 1996, 87, 173; Wrana et al., Trends Genet., 1996,12, 493-496. Smad proteins have been identified as signaling mediatorsof TGF-β super family. Hahn et al., Science, 1996, 271, 350-353. Geneticalterations in Smad2 and Smad4/DPC4 have been found in specific tumorsubsets, and thus Smads may function as tumor suppressor genes. Hahn etal., Science, 1996, 271, 350-353; Riggins et al., Nature Genet., 1996,13, 347-349; Eppert et al., Cell, 1996, 86, 543-552. Smad proteins sharetwo regions of high similarity, termed MH1 and MH2 domains, connectedwith a variable proline-rich sequence. Massague, Cell, 1996, 85,947-950; Derynck et al., Curr. Biol., 1996, 6, 1226-1229. The C-terminalpart of Smad2, when fused to a heterologous DNA-binding domain, wasfound to have transcriptional activity. Liu et al., Nature, 1996, 381,620-623; Meersseman et al., Mech. Dev., 1997, 61, 127-140. The intactSmad2 protein when fused to a DNA-binding domain, was latent, buttranscriptional activity was unmasked after stimulation with ligand. Liuet al., supra.

TGF-βinitiates an intracellular signaling pathway leading ultimately tothe expression of genes that regulate the cell cycle, controlproliferative responses, or relate to extracellular matrix proteins thatmediate outside-in cell signaling, cell adhesion, migration andintercellular communication.

TGF-β is also an important mediator of diabetic nephropathy, a commoncomplication in patients with either type 1 or type 2 diabetes mellitus.Ziyadeh et al., Proc. Natl. Acad. Sci., 2000, 97, 8015-8020 evaluatedthe role of renal TGF-β in the development of chronic structural andfunctional changes of diabetic nephropathy by assessing the response ofdb/db mice to chronic treatment with neutralizing anti-TGF-β1 andgeneralized (tubular and glomerular) up-regulation of TGF-β type IIreceptor. The antibody effectively prevented increases in renalexpression of matrix genes including type IV collagen and fibronectinand may have also stimulated matrix degradative pathways because TGF-βsuppresses the activity of metalloproteinases and increase theexpression of protease inhibitors such as plasminogen activatorinhibitor-1 (PAI-1).

There exists a need for effective therapeutic agents for inhibitingTGF-β activity, as well as for inhibiting the phosphorylation of smad2or smad3 by TGF-β type I or activin like kinase (ALK5) receptor and forpreventing and treating disease states mediated by the TGF-β signalingpathway in mammals. In particular, there continues to be a need forcompounds which selectively inhibit TGF-β.

SUMMARY OF THE INVENTION

The present invention provides compounds of the formula:

or a pharmaceutically acceptable salt or a prodrug thereof, wherein

-   -   Pg is hydrogen, alkyl, or a nitrogen protecting group;    -   n is an integer from 0 to 3;    -   each R¹ substituent is independently selected from the group        consisting of —R², -T-R², and -V-T-R;    -   wherein        -   each R² is independently selected from the group consisting            of C₁₋₃ aliphatic, hydroxy, —N(R³)₂, halo, cyano, —OR⁴,            —C(O)R⁴, —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴, —N(R³)C(O)R⁴,            —N(R³)CO₂R⁴, —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂,            —N(R³)C(O)N(R³)₂, —OC(O)R⁴, phenyl which is optionally            substituted with 1-3 R⁵, 5-6 membered heterocyclyl which is            optionally substituted with 1-3 R⁵, and 5-6 membered            heteroaryl which is optionally substituted with 1-3 R⁵;        -   each T is independently C₁₋₅ alkylidene that is optionally            interrupted by —O—, —C(O)—, —S—, —S(O)—, —S(O)₂—, or            —N(R³)—;        -   each V is independently selected from the group consisting            of —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —N(R³)—, —N(R³)C(O)—,            or —N(R³)C(O)₂—, —N(R³)S(O)₂—, —C(O)N(R³)—, —S(O)₂N(R³)—,            —N(R³)C(O)N(R³)—, and —OC(O)—;        -   each R³ is independently selected from the group consisting            of hydrogen, C₁₋₆ aliphatic, —C(O)R⁴, —C(O)₂R⁴, and —SO₂R⁴,        -   or two R³ on the same nitrogen together with their            intervening nitrogen form a 5-6 membered heterocyclyl or            heteroaryl ring having 1-3 ring heteroatoms selected from            nitrogen, oxygen or sulfur;        -   each R⁴ is independently selected from the group consisting            of C₁₋₆ aliphatic, phenyl or a 5-6 membered heteroaryl or            heterocyclyl having 1-3 ring heteroatoms selected from            nitrogen, oxygen or sulfur;    -   R⁸ is selected from the group consisting of:        -   (a) aryl, heteroaryl, and cycloalkyl, each of which is            optionally substituted with 1-3 R⁵, and        -   (b) alkyl;    -   wherein        -   each R⁵ is independently selected from the group consisting            of C₁₋₆ aliphatic, halo, —OH, —N(R³)₂, cyano, —OR⁴, —C(O)R⁴,            —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴, —N(R³)C(O)R⁴, —N(R³)CO₂R⁴,            —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂, —N(R³)C(O)N(R³)₂,            —OC(O)R⁴, —OC(O)N(R³)₂, phenyl, 5-6 membered heterocyclyl            and 5-6 membered heteroaryl,        -   or two adjacent R⁵ on an aryl, cycloalkyl, or heteroaryl            ring are taken together with their intervening atoms to form            a 5-6 membered fused ring having 0-2 heteroatoms selected            from nitrogen, oxygen or sulfur;    -   each R⁶ is independently selected from the group consisting of        hydrogen, halo, C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄ monoalkylamino and        C₁₋₄ dialkylamino; and    -   R⁷ is selected from the group consisting of hydrogen, halo, —OH,        —N(R³)₂, cyano, —OR⁴, —C(O)R⁴, —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴,        —N(R³)C(O)R⁴, —N(R³)CO₂R⁴, —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂,        —N(R³)C(O)N(R³)₂, and —OC(O)R⁴.

Compounds of Formula I, and compositions comprising the same, are usefulin a variety of pharmaceutical applications. In particular, compounds ofFormula I are useful in modulating TGF-β.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

In accordance with the present invention and as used herein, thefollowing terms are defined with the following meanings, unless statedotherwise.

“Alkyl” refers to a linear or branched saturated monovalent hydrocarbonmoiety having from one to twelve, preferably one to nine and morepreferably one to six, carbon atoms. Exemplary alkyl groups includemethyl, ethyl, n-propyl, 2-propyl, tert-butyl, pentyl, and the like.

The term “aliphatic” as used herein means straight-chain, branched orcyclic C₁-C₂ hydrocarbons which are completely saturated or whichcontain one or more units of unsaturation but which are not aromatic.For example, suitable aliphatic groups include substituted orunsubstituted linear, branched or cycloalkyl, alkenyl, alkynyl groupsand hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or(cycloalkyl)alkenyl. Preferred aliphatic groups are optionallysubstituted linear or branched alkyl, alkenyl, and alkynyl groups, withthe alkyl group being a particularly preferred aliphatic group.

An aliphatic group or a heterocyclic ring may contain one or moresubstituents. Examples of suitable substituents on the saturated carbonof an aliphatic group or of a heterocyclic ring include those listedherein for the unsaturated carbon of an aryl or heteroaryl group and thefollowing: ═O, ═S, ═NNHR*, ═NN(R*)₂, ═N—, ═NNHC(O)R*, ═NNHCO₂(alkyl),═NNHSO₂(alkyl), or ═NR*, where each R* is independently selected fromhydrogen, an unsubstituted aliphatic group and a substituted aliphaticgroup. Examples of substituents on the aliphatic group include amino,alkylamino, dialkylamino, aminocarbonyl, halogen, alkyl,alkylaminocarbonyl, dialkylaminocarbonyl, alkylaminocarbonyloxy,dialkylaminocarbonyloxy, alkoxy, nitro, cyano, carboxy, alkoxycarbonyl,alkylcarbonyl, hydroxy, haloalkoxy, and haloalkyl.

“Alkylidene” refers to an optionally substituted, straight or brancheddivalent carbon chain that may be fully saturated or have one or moreunits of unsaturation, e.g., alkylene, alkenylene, and alkynylene. Theoptional substituents are as described herein for an aliphatic group.Preferred alkylidene group is alkylene.

“Alkoxy” refers to a moiety of the formula —OR^(a), wherein R^(a) isalkyl as defined herein.

“Alkenyl” refers to a monovalent straight or branched chain containingtwo to twelve carbon atoms having one or more carbon-carbon doublebonds.

“Alkynyl” refers to a monovalent straight or branched chain containingtwo to twelve carbon atoms having one or more carbon-carbon triplebonds.

The term “cycloalkyl, used alone or as part of a larger moiety, includesmono- or bicyclic C₃-C₁₂ hydrocarbons which are completely saturated orwhich contain one or more units of unsaturation, but which are notaromatic. Exemplary cycloalkyl groups include, cyclopropyl, cyclopentyl,cyclohexyl, and the like.

“Aryl” refers to a monovalent mono-, bi- or tri-cyclic aromatichydrocarbon moiety having from 6 to 14 carbon ring atoms. Exemplary arylgroups include, but are not limited to, phenyl, 1-naphthyl, 2-naphthyl,anthracyl, and the like. Aryl group can be optionally substituted. Arylalso includes a group in which an aromatic ring is fused to one or morenon-aromatic rings, such as in an indanyl, phenanthridinyl, ortetrahydronaphthyl, where the radical or point of attachment is on thearomatic ring.

Terms “halo”, “halide” and “halogen” are used interchangeably herein andrefer to fluoro, chloro, bromo, or iodo.

The terms “haloalkyl”, “haloalkenyl” and “haloalkoxy”, mean alkyl,alkenyl or alkoxy, as the case may be, substituted with one or morehalogen atoms.

An aryl or heteroaryl group may contain one or more substituents.Examples of suitable substituents on the unsaturated carbon atom of anaryl and heteroaryl groups include a halogen, —R^(o), —OR^(o), —SR^(o),1,2-methylene-dioxy, 1,2-ethylenedioxy, protected OH (such as acyloxy),phenyl (Ph), substituted Ph, —O(Ph), substituted —O(Ph), —CH₂(Ph),substituted —CH₂(Ph), —CH₂CH₂(Ph), substituted —CH₂CH₂(Ph), —NO₂, —CN,—N(R^(o))₂, —NR^(o)C(O)R^(o), —NR^(o)C(O)N(R^(o))₂, —NR^(o)CO₂R^(o),—NR^(o)NR^(o)C(O)R^(o), —NR^(o)NR^(o)C(O)N(R^(o))₂,—NR^(o)NR^(o)CO₂R^(o), —C(O)C(O)R^(o), —C(O)CH₂C(O)R^(o), —CO₂R^(o),—C(O)R^(o), —C(O)N(R^(o))₂, —OC(O)N(R^(o))₂, —S(O)₂R^(o), —SO₂N(R^(o))₂,—S(O)R^(o), —NR^(o)SO₂N(R^(o))₂, —NR^(o)SO₂R^(o), —C(═S)N(R^(o))₂,—C(═NH)—N(R^(o))₂, —(CH₂)_(y)NHC(O)R^(o),—(CH₂)_(y)NHC(O)CH(V—R^(o))(R^(o)); wherein each R^(o) is independentlyselected from hydrogen, a substituted or unsubstituted aliphatic group,an unsubstituted heteroaryl and heterocyclic ring, phenyl (Ph),substituted Ph, —O(Ph), substituted —O(Ph), —CH₂(Ph), and substituted—CH₂(Ph); wherein y is 0-6; and V is a linker group. Examples ofsubstituents on the aliphatic group or the phenyl ring of R^(o) includeamino, alkylamino, dialkylamino, aminocarbonyl, halogen, alkyl,alkylaminocarbonyl, dialkylaminocarbonyl, alkylaminocarbonyloxy,dialkylaminocarbonyloxy, alkoxy, nitro, cyano, carboxy, alkoxycarbonyl,alkylcarbonyl, hydroxy, haloalkoxy, and haloalkyl.

The term “linker group” or “linker” means an organic moiety thatconnects two parts of a compound. Linkers are typically comprised of anatom such as oxygen or sulfur, a unit such as —NH—, —CH₂—, —C(O)—,—C(O)NH—, or a chain of atoms, such as an alkylidene chain. Themolecular mass of a linker is typically in the range of about 14 to 200,preferably in the range of 14 to 96 with a length of up to about sixatoms. Examples of linkers include a saturated or unsaturated C₁₋₆alkylidene chain which is optionally substituted, and wherein one or twosaturated carbons of the chain are optionally replaced by —C(O)—,—C(O)C(O)—, —CONH—, —CONHNH—, —CO₂—, —OC(O)—, —NHCO₂—, —O—, —NHCONH—,—OC(O)NH—, —NHNH—, —NHCO—, —S—, —SO—, —SO₂—, —NH—, —SO₂NH—, or —NHSO₂—.—“Haloalkyl” refers to an alkyl group as defined herein that issubstituted with one or more same or different halo atoms, e.g., —CH₂Cl,—CF₃, —CH₂CF₃, —CH₂CCl₃, and the like. The term “haloalkyl” alsoincludes those alkyl groups in which all alkyl hydrogen atoms arereplaced by halogen atoms, such as perfluoroalkyl groups.

“Heteroaryl” refers to a monovalent mono-, bi- or tri-cyclic aromaticmoiety of five to twenty, preferably five to fourteen, ring atomscontaining one to four, preferably one to three, hetero ring atoms eachof which is independently selected from the group N, O, and S(O)_(z)(wherein the subscript z is 0, 1 or 2), the remaining ring atoms beingC. Exemplary heteroaryl groups include, but are not limited to,2-furanyl, 3-furanyl, 3-furazanyl, N-imidazolyl, 2-imidazolyl,4-imidazolyl, 5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl,2-oxadiazolyl, 5-oxadiazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl,1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 1-pyrazolyl, 2-pyrazolyl,3-pyrazolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl,5-pyrimidyl, 3-pyridazinyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl,5-tetrazolyl, 2-triazolyl, 5-triazolyl, 2-thienyl, 3-thienyl,carbazolyl, benzimidazolyl, benzothienyl, benzofuranyl, indolyl,quinolinyl, benzotriazolyl, benzothiazolyl, benzooxazolyl,benzimidazolyl, isoquinolinyl, indazolyl, isoindolyl, acridinyl,benzoisoxazolyl, and the like. Heteroaryl also includes a group in whicha heteroatomic ring is fused to one or more aromatic or nonaromaticrings where the point of attachment is on the heteroaromatic ring.Examples include tetrahydroquinolinyl, tetrahydroisoquinolinyl, andpyrido[3,4-d]pyrimidinyl. Heteroaryl can be optionally substituted.

“Heterocyclyl” refers to a non-aromatic mono-, bi- or tri-cyclic moietyhaving three to twenty, preferably five to fourteen, ring atoms in whichone to four, preferably one to three, and more preferably one or two,ring atoms are heteroatoms each of which is independently selected fromN, O, and S(O)_(z) (wherein the subscript z is 0, 1 or 2), the remainingring atoms being C, with the understanding that the number of heteroring atoms is less than the total number of ring atoms present in theheterocyclyl moiety. Exemplary heterocyclyl groups include, but are notlimited to, piperidinyl, piperazinyl, morpholino, pyrrolidinyl,3-1H-benzimidazol-2-one, (1-substituted)-2-oxo-benzimidazol-3-yl,2-tetrahydrofuranyl, 3-tetrahydrofuranyl, 2-tetrahydropyranyl,3-tetrahydropyranyl, 4-tetrahydropyranyl, [1,3]-dioxalanyl,[1,3]-dithiolanyl, [1,3]-dioxanyl, 2-tetrahydrothiophenyl,3-tetrahydrothiophenyl, 2-morpholinyl, 3-morpholinyl, 4-morpholinyl,2-thiomorpholinyl, 3-thiomorpholinyl, 4-thiomorpholinyl, 1-pyrrolidinyl,2-pyrrolidinyl, 3-pyrrolidinyl, 1-piperazinyl, 2-piperazinyl,1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl,4-thiazolidinyl, diazolonyl, N-substituted diazolonyl, 1-phthalimidinyl,benzoxanyl, benzopyrrolidinyl, benzopiperidinyl, benzoxolanyl,benzothiolanyl, and benzothianyl. Also included within the scope of theterm “heterocyclyl” is a group in which a non-aromaticheteroatom-containing ring is fused to one or more aromatic ornon-aromatic rings, such as in an indolinyl, chromanyl, phenanthridinyl,or tetrahydroquinolinyl, where the radical or point of attachment is onthe non-aromatic heteroatom-containing ring. Heterocyclyl can also beoptionally substituted.

“Pharmaceutically acceptable excipient (or carrier)” means an excipientthat is useful in preparing a pharmaceutical composition that isgenerally safe. Suitable pharmaceutically acceptable excipients are wellknown to one skilled in the art and include excipients that areacceptable for veterinary or human pharmaceutical use.

“Protecting group” refers to a moiety, except alkyl groups, that whenattached to a reactive group in a molecule masks, reduces or preventsthat reactivity. Examples of protecting groups can be found in T. W.Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3^(rd)edition, John Wiley & Sons, New York, 1999, and Harrison and Harrison etal., Compendium of Synthetic Organic Methods, Vols. 1-8 (John Wiley andSons, 1971-1996), which are incorporated herein by reference in theirentirety. Representative hydroxy protecting groups include acyl groups,benzyl and trityl ethers, tetrahydropyranyl ethers, trialkylsilyl ethersand allyl ethers. Representative amino protecting groups include,formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl (CBZ),tert-butoxycarbonyl (Boc), trimethyl silyl (TMS),2-trimethylsilyl-ethanesulfonyl (SES), trityl and substituted tritylgroups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl (FMOC),nitro-veratryloxycarbonyl (NVOC), and the like.

“Leaving group” has the meaning conventionally associated with it insynthetic organic chemistry, i.e., an atom or a group capable of beingdisplaced by a nucleophile and includes halo (such as chloro, bromo, andiodo), alkanesulfonyloxy, arenesulfonyloxy, alkylcarbonyloxy (e.g.,acetoxy), arylcarbonyloxy, mesyloxy, tosyloxy,trifluoromethanesulfonyloxy, aryloxy (e.g., 2,4-dinitrophenoxy),methoxy, N,O-dimethylhydroxylamino, and the like.

“Pharmaceutically acceptable salt” of a compound means a salt that ispharmaceutically acceptable and that possesses the desiredpharmacological activity of the parent compound. Such salts include: (1)acid addition salts, formed with inorganic acids such as hydrochloricacid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, andthe like; or formed with organic acids such as acetic acid, propionicacid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvicacid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid,3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like; or (2)salts formed when an acidic proton present in the parent compound eitheris replaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, tromethamine,N-methylglucamine, and the like.

The terms “pro-drug” and “prodrug” are used interchangeably herein andrefer to any compound which releases an active parent drug according toFormula I in vivo when such prodrug is administered to a mammaliansubject. Prodrugs of a compound of Formula I are typically prepared bymodifying one or more functional group(s) present in the compound ofFormula I in such a way that the modification(s) may be cleaved in vivoto release the parent compound. Prodrugs include compounds of Formula Iwherein a hydroxy, amino, or sulfhydryl group in a compound of Formula Iis bonded to any group (e.g., protecting group) that may be cleaved invivo to regenerate the free hydroxyl, amino, or sulfhydryl group,respectively. Examples of prodrugs include, but are not limited to,esters (e.g., acetate, formate, and benzoate derivatives) and carbamates(e.g., N,N-dimethylaminocarbonyl) of hydroxy functional groups, andamides, carbamates and urea derivatives of amino functional groups, andthe like. Prodrug forms often offer advantages of solubility, tissuecompatibility, or delayed release in the mammalian organism (see,Bundgard, Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985and Silverman, The Organic Chemistry of Drug Design and Drug Action, pp.352-401, Academic Press, San Diego, Calif., 1992, all of which areincorporated herein by reference in their entirety). Moreover, theprodrug derivatives of the invention may be combined with other featuresknown to one skilled in the art to enhance bioavailability.

As used herein, the terms “treating”, “contacting” and “reacting”, whenreferring to a chemical reaction, are used interchangeably herein andrefer to adding or mixing two or more reagents under appropriateconditions to produce the indicated and/or the desired product. Itshould be appreciated that the reaction which produces the indicatedand/or the desired product may not necessarily result directly from thecombination of two or more reagents which were initially added, i.e.,there may be one or more intermediates which are produced in the mixturewhich ultimately leads to the formation of the indicated and/or thedesired product.

“Treating” or “treatment” of a disease includes: (1) preventing thedisease, i.e., causing the clinical symptoms of the disease not todevelop in a mammal that may be exposed to or predisposed to the diseasebut does not yet experience or display symptoms of the disease; (2)inhibiting the disease, i.e., arresting or reducing the development ofthe disease or its clinical symptoms; or (3) relieving the disease,i.e., causing regression of the disease or its clinical symptoms.

“A therapeutically effective amount” means the amount of a compoundthat, when administered to a subject or patient for treating a disease,is sufficient to effect such treatment for the disease. The“therapeutically effective amount” will vary depending on a variety ofthe factors, such as a particular compound used, the disease and itsseverity, the age, weight, and other factors of the subject to betreated.

As used herein, the terms “as defined herein” and “as defined above”when referring to a variable incorporates by reference the broaddefinition of the variable as well as preferred, more preferred and mostpreferred definitions, if any.

“Biological property” for the purposes herein means an in vivo effectoror activity that is directly or indirectly performed by a compound thatare often shown by in vitro assays. Effector functions include receptoror ligand binding, any enzyme activity or enzyme modulatory activity,any carrier binding activity, any hormonal activity, any activity inpromoting or inhibiting adhesion of cells to an extracellular matrix orcell surface molecules, or any structural role.

A combination of substituents or variables is permissible only if such acombination results in a stable or chemically feasible compound. Astable compound or chemically feasible compound is one in which thechemical structure is not substantially altered when kept at atemperature of 40° C. or less, in the absence of moisture or otherchemically reactive conditions, for at least a week.

Unless otherwise stated, structures depicted herein are also meant toinclude all stereochemical forms of the structure; i.e., the R and Sconfigurations for each asymmetric center. Therefore, singlestereochemical isomers as well as enantiomeric and diastereomericmixtures of the present compounds are within the scope of the invention.Unless otherwise stated, structures depicted herein are also meant toinclude compounds which differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures except for the replacement of a hydrogen atom by a deuteriumor tritium, or the replacement of a carbon by ¹³C or ¹⁴C are within thescope of this invention.

Compounds of the Present Invention

In one aspect, the present invention provides compounds of the formula:

wherein

-   -   Pg is hydrogen, alkyl, or a nitrogen protecting group;    -   n is an integer from 0 to 3;    -   each R¹ substituent is independently selected from the group        consisting of —R², -T-R², and -V-T-R;    -   wherein        -   each R² is independently selected from the group consisting            of C₁₋₃ aliphatic, hydroxy, —N(R³)₂, halo, cyano, —OR⁴,            —C(O)R⁴, —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴, —N(R³)C(O)R⁴,            —N(R³)CO₂R⁴, —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂,            —N(R³)C(O)N(R³)₂, —OC(O)R⁴, phenyl which is optionally            substituted with 1-3 R⁵, 5-6 membered heterocyclyl which is            optionally substituted with 1-3 R⁵, and 5-6 membered            heteroaryl which is optionally substituted with 1-3 R⁵;        -   each T is independently C₁₋₅ alkylidene that is optionally            interrupted by —O—, —C(O)—, —S—, —S(O)—, —S(O)₂—, or            —N(R³)—;        -   each V is independently selected from the group consisting            of —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —N(R³)—, —N(R³)C(O)—,            or —N(R³)C(O)₂—, —N(R³)S(O)₂—, —C(O)N(R³)—, —S(O)₂N(R³)—,            —N(R³)C(O)N(R³)—, and —OC(O)—;        -   each R³ is independently selected from the group consisting            of hydrogen, C₁₋₆ aliphatic, —C(O)R⁴, —C(O)₂R⁴, and —SO₂R⁴,        -   or two R³ on the same nitrogen together with their            intervening nitrogen form a 5-6 membered heterocyclyl or            heteroaryl ring having 1-3 ring heteroatoms selected from            nitrogen, oxygen or sulfur;        -   each R⁴ is independently selected from the group consisting            of C₁₋₆ aliphatic, phenyl or a 5-6 membered heteroaryl or            heterocyclyl having 1-3 ring heteroatoms selected from            nitrogen, oxygen or sulfur;    -   R⁸ is selected from the group consisting of:        -   (a) aryl, heteroaryl, and cycloalkyl, each of which is            optionally substituted with 1-3 R⁵, and        -   (b) alkyl;    -   wherein        -   each R⁵ is independently selected from the group consisting            of C₁₋₆ aliphatic, halo, —OH, —N(R³)₂, cyano, —OR⁴, —C(O)R⁴,            —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴, —N(R³)C(O)R⁴, —N(R³)CO₂R⁴,            —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂, —N(R³)C(O)N(R³)₂,            —OC(O)R⁴, —OC(O)N(R³)₂, phenyl, 5-6 membered heterocyclyl            and 5-6 membered heteroaryl,        -   or two adjacent R⁵ on an aryl, cycloalkyl, or heteroaryl            ring are taken together with their intervening atoms to form            a 5-6 membered fused ring having 0-2 heteroatoms selected            from nitrogen, oxygen or sulfur;    -   each R⁶ is independently selected from the group consisting of        hydrogen, halo, C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄ monoalkylamino and        C₁₋₄ dialkylamino; and    -   R⁷ is selected from the group consisting of hydrogen, halo, —OH,        —N(R³)₂, cyano, —OR⁴, —C(O)R⁴, —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴,        —N(R³)C(O)R⁴, —N(R³)CO₂R⁴, —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂,        —N(R³)C(O)N(R³)₂, and —OC(O)R⁴.

The compounds of the present invention can exist in unsolvated forms aswell as solvated forms, including hydrated forms, all of which areintended to be encompassed within the scope of the invention. The scopeof the present invention also includes all pharmaceutically acceptablesalts, prodrugs and isomers, particularly stereoisomers, of compounds ofFormula I. Unless otherwise stated, the term “stereoisomer” includes apure chiral form, a racemic mixture and an enantiomerically and/ordiastereomerically enriched forms. Furthermore, some compounds of thepresent invention can exist in tautomeric forms which are also intendedto be encompassed within the scope of the present invention.

Some compounds of Formula I are capable of forming pharmaceuticallyacceptable salts. As stated above, these salts are also contemplated tobe within the scope of the present invention. Pharmaceuticallyacceptable acid addition salts of the compounds of Formula I includesalts derived from inorganic acids such as hydrochloric, nitric,phosphoric, sulfuric, hydrobromic, hydriodic, phosphorus, and the like,as well as the salts derived from organic acids, such as aliphatic mono-and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, alkanedioic acids, aromatic acids, aliphatic andaromatic sulfonic acids, etc. Such salts include sulfate, pyrosulfate,bisulfate, sulfite, bisulfite, nitrate, phosphate,monohydrogenphosphate, dihydrogenphosphate, metaphosphate,pyrophosphate, chloride, bromide, iodide, acetate, propionate,caprylate, isobutyrate, oxalate, malonate, succinate, suberate,sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate,methylbenzoate, dinitrobenzoate, pthalate, benzenesulfonate,toluenesulfonate, phenylacetate, citrate, lactate, maleate, tartarate,methanesulfonate, and the like. Also contemplated are salts of aminoacids, such as arginate, gluconate and galacturonate salts and the like(see, for example, Berge et al., “Pharmaceutical Salts,” J. ofPharmaceutical Science, 1977, 66, 1-19, which is incorporated herein byreference in its entirety).

The acid addition salts of compounds of Formula I can be prepared bycontacting the free base form with a sufficient amount of the desiredacid to produce the salt in the conventional manner. The free base formcan be regenerated by contacting the salt form with a base and isolatingthe free base in the conventional manner. The free base forms may differfrom their respective salt forms somewhat in certain physical propertiessuch as solubility in polar solvents, but otherwise the salts aresubstantially equivalent to their respective free base for the purposesof the present invention.

Pharmaceutically acceptable base addition salts of compounds of FormulaI can be formed with metal ions or amines, such as alkali and alkalineearth metal ions or organic amines. Examples of metal ions which areused as cations include sodium, potassium, magnesium, calcium, and thelike. Examples of suitable amines are N, N′-dibenzylethylenediamine,chlorocaine, choline, diethanolamine, ethylenediamine,N-methylglucamine, and procaine (see, for example, Berge et al., supra).

The base addition salts of acidic compounds can be prepared bycontacting the free acid form with a sufficient amount of the desiredbase to produce the salt in the conventional manner. The free acid formcan be regenerated by contacting the salt form with an acid andisolating the free acid in the conventional manner. The free acid formsmay differ from their respective salt forms somewhat in certain physicalproperties such as solubility in polar solvents, but otherwise the saltsare substantially equivalent to their respective free acid for thepurposes of the present invention.

In one embodiment, compounds of the present invention are of theformula:

In another embodiment, compounds of the present invention are of theformula:

Still in another embodiment, R⁸ in compounds of Formulas I, IA and IB isselected from phenyl, naphthyl, pyridyl, thienyl, furyl (each of whichis optionally substituted with 1-3 R⁵), cyclohexyl, cyclopentyl,cyclopropyl, and t-butyl. When R⁸ is a mono-substituted phenyl, thepreferred substituent position on the phenyl ring is meta-positionrelative to the quinoline ring system. A particularly preferred R⁸ isselected from 2-fluorophenyl, benzo[1,3]dioxol-5-yl,2-trifluoromethylphenyl, 2-chlorophenyl, pyridin-4-yl, 2-methylphenyl,2-methoxyphenyl, 3-fluorophenyl, 3-chlorophenyl, 5-chloro-thiophen-2-yl,5-chloro-furan-2-yl, 5-methyl-thiophen-2-yl, 5-methyl-furan-2-yl,4-methyl-thiophen-2-yl, 2,3-dihydro-benzofuran-5-yl,2-methylsulfanylphenyl, 4-fluorophenyl, 2-methanesulfinylphenyl,4-methoxyphenyl, 2-cyanophenyl, 2-amidophenyl, 4-hydroxyphenyl,3-aminophenyl, 3-methoxyphenyl, 4-cyanophenyl, 2,6-dichlorophenyl,phenyl, 4-chlorophenyl, cyclohexyl, cyclopropyl, cyclopentyl,4-methylsulfanylphenyl, t-butyl, 4-amidophenyl, naphthalen-2-yl,4-methanesulfinylphenyl, and 2-bromophenyl.

In another embodiment, R⁸ in compounds of Formulas I, IA and IB is a 5-6membered aryl or heteroaryl selected from phenyl, pyridyl, thienyl, andfuryl, each of which is optionally substituted with 1-3 R⁵.

Yet in another embodiment, each R¹ in compounds of Formulas I, IA and IBis independently selected from thienyl, furyl, pyrrolyl, and phenyl,each of which is optionally substituted with 1-3 R⁵; halo; —OR⁴;—N(R³)₂; and —N(R³)-T-OR⁴. Particularly preferred R¹ are those shown inrepresentative compounds of Formula I below, for example, methylamino,2-methoxyethylamino, furanyl, thiophenyl (i.e., thienyl), phenyloptionally substituted with 1-3 R⁵, halogen and methoxy. An especiallypreferred R¹ is independently selected from methylamino,2-methoxyethylamino, furan-2-yl, thiophen-3-yl, phenyl optionallysubstituted with 1-2 R⁵, bromo and methoxy.

Still further, combinations of the different embodiments and preferredgroups described herein will form other embodiments and preferredgroups. For example, one particularly preferred group of compounds ofFormula I are those represented by Formula IA in which R¹ is selectedfrom methylamino, 2-methoxyethylamino, optionally substituted phenyl,furan-2-yl and thiophen-3-yl. Still in another particular embodiment, apreferred group of compounds of Formula I are those represented byFormula IB in which R¹ is selected from methoxy, methylamino,2-methoxyethylamino and bromo. In this manner, a variety of preferredcompounds are embodied within the present invention. Some of therepresentative compounds of Formula I are shown in Table 1 below. TABLE1 Representative compounds of Formula I. (1)

[3-Fluoro-2-(2-methylsulfanyl-phenyl)-quinolin-4-yl]- pyridin-4-yl-amine(2)

[3-Fluoro-2-(2-methanesulfinyl-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine (3)

2-[3-Fluoro-4-(pyridin-4-ylamino)-quinolin-2-yl]- benzonitrile (4)

2-[3-Fluoro-4-(pyridin-4-ylamino)- quinolin-2-yl]-benzamide (5)

[3-Fluoro-2-(4-fluoro-phenyl)-quinolin-4-yl]- pyridin-4-yl-amine (6)

[3-Fluoro-2-(4-methoxy-phenyl)-quinolin-4-yl]- pyridin-4-yl-amine (7)

[2-(3-Chloro-phenyl)-3-fluoro-quinolin-4-yl]- pyridin-4-yl-amine (8)

[3-Fluoro-2-(3-fluoro-phenyl)-quinolin-4-yl]- pyridin-4-yl-amine (9)

4-[3-Fluoro-4-(pyridin-4-ylamino)- quinolin-2-yl]-phenol (10)

[3-Fluoro-2-(3-methoxy-phenyl)- quinolin-4-yl]-pyridin-4-yl-amine (11)

[2-(2,6-Dichloro-phenyl)-3-fluoro- quinolin-4-yl]-pyridin-4-yl-amine(12)

[2-(4-Chloro-phenyl)-3-fluoro- quinolin-4-yl]-pyridin-4-yl-amine (13)

[2-(3-Amino-phenyl)-3-fluoro-quinolin-4-yl]- pyridin-4-yl-amine (14)

4-[3-Fluoro-4-(pyridin-4-ylamino)-quinolin-2-yl]- benzonitrile (15)

(3-Fluoro-2-phenyl-quinolin-4-yl)- pyridin-4-yl-amine (16)

(2-Cyclohexyl-3-fluoro-quinolin-4-yl)- pyridin-4-yl-amine (17)

(2-Cyclopropyl-3-fluoro- quinolin-4-yl)-pyridin-4-yl-amine (18)

(2-tert-Butyl-3-fluoro-quinolin-4-yl)- pyridin-4-yl-amine (19)

(3-Fluoro-2-naphthalen-2-yl- quinolin-4-yl)-pyridin-4-yl-amine (20)

(2-Cyclopentyl-3-fluoro-quinolin-4-yl)- pyridin-4-yl-amine (21)

[3-Fluoro-2-(4-methylsulfanyl-phenyl)- quinolin-4-yl]-pyridin-4-yl-amine(22)

4-[3-Fluoro-4-(pyridin-4-ylamino)- quinolin-2-yl]-benzamide (23)

(3-Fluoro-2-phenyl-quinolin-4-yl)- pyridin-4-yl-amine (24)

[3-Fluoro-2-(4-methanesulfinyl-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine (25)

3-Fluoro-2-pyridin-4-yl-4- pyrrolo[3,2-c]pyridin-1-yl-quinoline (26)

3-(3-Fluoro-4-pyrrolo[3,2-c]pyridin-1-yl- quinolin-2-yl)-phenylamine(27)

[3-Fluoro-2-(2-fluoro-phenyl)- quinolin-4-yl]-pyridin-4-yl-amine (28)

[2-(2-Bromo-phenyl)-3-fluoro- quinolin-4-yl]-pyridin-4-yl-amine (29)

(2-Benzo[1,3]dioxol-5-yl-3-fluoro- quinolin-4-yl)-pyridin-4-yl-amine(30)

[3-Fluoro-2-(2-trifluoromethyl-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine (31)

(3-Fluoro-2-pyridin-4-yl-quinolin-4-yl)- pyridin-4-yl-amine (32)

[2-(2-Chloro-phenyl)-3-fluoro- quinolin-4-yl]-pyridin-4-yl-amine (33)

(3-Fluoro-2-o-tolyl-quinolin-4-yl)- pyridin-4-yl-amine (34)

[3-Fluoro-2-(2-methoxy-phenyl)- quinolin-4-yl]-pyridin-4-yl-amine (35)

3-Fluoro-2-(2-fluoro-phenyl)-N⁶-methyl-N⁴-pyridin-4-yl-quinoline-4,6-diamine (36)

2-Benzo[1,3]dioxol-5-yl-3-fluoro-N⁶-methyl-N⁴-pyridin-4-yl-quinoline-4,6-diamine (37)

3-Fluoro-2-(2-fluoro-phenyl)-N⁶-(2-methoxy-ethyl)-N⁴-pyridin-4-yl-quinoline-4,6-diamine (38)

2-Benzo[1,3]dioxol-5-yl-3-fluoro-N⁶-(2-methoxy-ethyl)-N⁴-pyridin-4-yl-quinoline-4,6-diamine (39)

3-[3-Fluoro-2-(2-fluoro-phenyl)-4-(pyridin-4-ylamino)-quinolin-6-yl]-benzamide (40)

X = S, O, NH When X = O, [3-Fluoro-2-(2-fluoro-phenyl)-6-furan-2-yl-quinolin-4-yl]-pyridin-4-yl-amine (41)

X = S, O, NH When X = S, [3-Fluoro-2-(2-fluoro-phenyl)-6-thiophen-3-yl-quinolin-4-yl]-pyridin-4-yl-amine (42)

R⁹ = C₁₋₃ alkyl, halo, —OH, N(R³)₂, cyano, or —OR⁴, where R³ and R⁴ areas defined herein (43)

(2-Benzo[1,3]dioxol-5-yl-7-bromo-3-fluoro-quinolin-4-yl)-pyridin-4-yl-amine (44)

[3-Fluoro-2-(2-fluoro-phenyl)-7-methoxy-quinolin-4-yl]-pyridin-4-yl-amine (45)

2-Benzo[1,3]dioxol-5-yl-3-fluoro-N⁷-methyl-N⁴-pyridin-4-yl-quinoline-4,7-diamine (46)

3-Fluoro-2-(2-fluoro-phenyl)-N⁷-methyl-N⁴-pyridin-4-yl-quinoline-4,7-diamine (47)

2-Benzo[1,3]dioxol-5-yl-3-fluoro-N⁷-(2-methoxy-ethyl)-N⁴-pyridin-4-yl-quinoline-4,7-diamine (48)

R¹⁰ is independently selected from thienyl, furyl, pyrrolyl, and phenyl,each of which is optionally substituted with 1-3 R⁵; halo; —OR⁴;—N(R³)₂; and —N(R³)—T—OR⁴, where R³, R⁴, R⁵ and T are as defined herein.Particularly preferred R¹⁰ is methylamino, 2- methoxyethylamino,furanyl, thiophenyl (i.e., thienyl), phenyl optionally substituted with1-3 R⁵, halogen or methoxy. (49)

(2-Benzo[1,3]dioxol-5-yl-3-fluoro-7-methoxy-quinolin-4-yl)-pyridin-4-yl-amine, (50)

[3-Fluoro-2-(3-fluoro-phenyl)- quinolin-4-yl]-pyridin-4-yl-amine (51)

[2-(5-Chloro-thiophen-2-yl)-3- fluoro-quinolin-4-yl]-pyridin-4-yl-amine(52)

[2-(3-Chloro-phenyl)-3-fluoro- quinolin-4-yl]-pyridin-4-yl-amine (53)

[2-(5-Chloro-furan-2-yl)-3-fluoro- quinolin-4-yl]-pyridin-4-yl-amine(54)

[3-Fluoro-2-(5-methyl-thiophen-2-yl)- quinolin-4-yl]-pyridin-4-yl-amine(55)

[3-Fluoro-2-(5-methyl-furan-2-yl)- quinolin-4-yl]-pyridin-4-yl-amine(56)

[3-Fluoro-2-(4-methyl-thiophen-2-yl)- quinolin-4-yl]-pyridin-4-yl-amine(57)

[2-(2,3-Dihydro-benzofuran-5-yl)-3-fluoro-quinolin-4-yl]-pyridin-4-yl-amine (58)

where R¹¹ = alkyl, benzyl, or phenyl (59)

where R¹² = H, Me, or —CH₂CH₂-piperidine (60)

where R¹³ = alkyl, benzyl, phenyl or alkyloxy (61)

[2-(3-Bromo-phenyl)-3-fluoro- quinolin-4-yl]-pyridin-4-yl-amine (62)

(2-Chloro-pyridin-4-yl)-[3-fluoro-2-(5-methyl-furan-2-yl)-quinolin-4-yl]-amine (63)

(2-Chloro-pyridin-4-yl)-[2-(5-chloro-thiophen-2-yl)-3-fluoro-quinolin-4-yl]-amine (64)

(2-Chloro-pyridin-4-yl)-[3-fluoro-2-(3-fluoro-phenyl)-quinolin-4-yl]-amine (65)

[3-Fluoro-2-(5-methyl-furan-2-yl)-quinolin-4-yl]-methyl-pyridin-4-yl-amine and (66)

(2-Chloro-pyridin-4-yl)-[2-(5-chloro-thiophen-2-yl)-3-fluoro-quinolin-4-yl]-methyl-amineSynthesis

The compounds of the present invention may be synthesized by a varietyof methods known to one of ordinary skill in the art or depicted in theillustrative synthetic reaction schemes shown and described below.Starting materials used in any of these methods are commerciallyavailable from chemical vendors such as Aldrich, Sigma, NovaBiochemicals, Bachem Biosciences, and the like, or can be prepared bymethods known to those skilled in the art following procedures set forthin references such as Fieser and Fieser's Reagents for OrganicSynthesis; Wiley & Sons: New York, 1991, Volumes 1-15; Rodd's Chemistryof Carbon Compounds, Elsevier Science Publishers, 1989, Volumes 1-5 andSupplementals; and Organic Reactions, Wiley & Sons: New York, 1991,Volumes 1-40.

It should be appreciated that the following synthetic reaction schemesare merely illustrative of some methods by which the compounds of thepresent invention can be synthesized, and various modifications to thesesynthetic reaction schemes can be made and will be suggested to oneskilled in the art having referred to the disclosure contained in thisApplication.

Unless specified to the contrary, the reactions described hereinpreferably are conducted under an inert atmosphere at atmosphericpressure at a reaction temperature range of from about −78° C. to about150° C., more preferably from about 0° C. to about 125° C., and mostpreferably and conveniently at about room (or ambient) temperature,e.g., about 20° C.

The starting materials and the intermediates of the synthetic reactionschemes can be isolated and purified, if desired, using conventionaltechniques, including but not limited to, filtration, distillation,crystallization, chromatography, and the like. Such materials can becharacterized using conventional means, including physical constants andspectral data.

As shown in Scheme 1 above, 3-aminoquinoline was converted to3-fluoroquinoline. This conversion is typically achieved by convertingthe amino group to a diazonium salt and displacing the diazonium moietywith a fluoride ion. Lithiation of 3-fluoroquinoline with a strong base,e.g., lithium diisopropyl amide, followed by treatment with iodine gave3-fluoro-4-iodoquinoline. Deprotonation of 3-fluoro-4-iodoquinoline witha base, e.g., lithium diisopropyl amide, and reacting the deprotonatedmoiety with hexachloroethane gave 4-chloro-3-fluoro-2-iodoquinoline.Reacting 4-chloro-3-fluoro-2-iodoquinoline with a boronic acidderivative R³—B(OH)₂ gave the 2-substituted quinoline compound. A metalhalogen exchange reaction between the 2-substituted quinoline compoundand butyl lithium followed by quenching with iodine then gave a2-substituted-3-fluoro-3-iodoquinoline compound, which was subjected toa palladium mediated cross-coupling reaction with an amine compound,e.g., R¹—NH₂, to yield the desired compound.

As shown in Scheme 2, instead of using a palladium catalyst mediatedcross-coupling reaction, the iodo group on the 4-position of thequinoline compound can also be displaced with an amino compound. Forexample, reacting a 2-aryl-6-bromo-3-fluoro-4-iodoquinoline compoundwith 4-aminopyridine in the presence of a base gave a[2-aryl-6-bromo-3-fluoroquinolin-4-yl]pyridyl-4-yl amine compound, whichcan be further modified by reacting with a various amino compounds,arylboronic acids or heteroaryl boronic acids using a palladium mediatedcross-coupling reaction to afford the desired6-substituted-2-aryl-3-fluoroquinoline compounds.

Utility

Compounds of the present invention possess a wide variety of biologicalactivities. In particular, compounds of the present invention are usefulin modulating, especially inhibiting, TGF-β signaling pathway.Accordingly, compounds of the present invention are useful in treating apatient having a TGF-β mediated disorder. Treatment of such a patientgenerally involves administering a therapeutically effective amount of acompound of Formula I to the patient in need of such a treatment.

As used herein, a subject or a patient can be any mammal, so long as themammal is in need of modulation of a pathological or biological processmediated by compounds of the present invention. The term “mammal” isdefined as a species belonging to the class Mammalia. Examples ofmammals include mice, rats, cows, sheep, pigs, goats, horses, bears,monkeys, dogs, cats and, preferably, humans. Transgenic organisms whichexpress TGF-β are also included in this definition.

TGF-β is a potent regulator of the cell cycle in many cell typesincluding vascular smooth muscle (VSM) and endothelial cells and, as aresult, TGF-β is believed to play an important role in vascularproliferative process such as angiogenesis. Pathological conditionsassociated with VSM include restenosis, atherosclerosis, coronary heartdisease, thrombosis, myocardial infarction, stroke, smooth muscleneoplasms such as leiomyoma and leiomyosarcoma of the bowel and uterus,uterine fibroid or fibroma, and obliterative disease of vascular graftsand transplanted organs. Other vascular diseases include unstableangina, chronic stable angina, transient ischemic attacks, strokes,peripheral vascular disease, preeclampsia, deep venous thrombosis,embolism, disseminated intravascular coagulation or thrombotic cytopenicpurpura. Vascular injury includes an injury arising by any meansincluding, but not limited to, procedures such as angioplasty, carotidendarterectomy, post CABG (coronary artery bypass graft) surgery,vascular graft surgery, stent placements or insertion of endovasculardevices and prostheses.

Compounds of the present invention can be used to treat smooth musclecell proliferation. In one particular embodiment, a compound of thepresent invention is used to inhibit or reduce stenosis due toproliferation of vascular smooth muscle cells following traumatic injuryto vessels such as during vascular surgery. The present invention alsocontemplates the delivery of a compound of Formula I to vascular smoothmuscle cells to exert inhibitory effects over an extended period oftime.

The TGF-β inhibition activity is useful in treating fibroproliferativediseases, treating collagen vascular disorders, treating eye diseasesassociated with a fibroproliferative condition, venting excessivescarring, treating neurological conditions and other conditions that aretargets for TGF-β inhibitors and in preventing excessive scarring thatelicits and accompanies restenosis following coronary angioplasty,cardiac fibrosis occurring after infarction and progressive heartfailure, and in hypertensive vasculopathy, and keloid formation orhypertrophic scars occurring during the healing of wounds includingsurgical wounds and traumatic lacerations. Neurological conditionscharacterized by TGF-β production include CNS injury after traumatic andhypoxic insults, Alzheimer's disease, and Parkinson's disease.

Other conditions that are potential clinical targets for TGF-βinhibitors include myelofibrosis, tissue thickening resulting fromradiation treatment, nasal polyposis, polyp surgery, liver cirrhosis,and osteoporosis.

The phrase “TGF-β disease condition” includes those states, disorders,or diseases characterized by aberrant or undesirable activity orexpression of TGF-β. Examples of TGF-β associated disease conditionsinclude, but are not limited to, disorders involving or associated withcardiovascular diseases such as myocardial infarction, stroke,thrombosis, congestive heart failure, dilated cardiomyopathy,myocarditis, or vascular stenosis associated with atherosclerosis,angioplasty treatment, or surgical incisions or mechanical trauma;kidney diseases associated with fibrosis and/or sclerosis, includingglomerulonephritis of all etiologies, diabetic nephropathy, and allcauses of renal interstitial fibrosis, including hypertension,complications of drug exposure, such as cyclosporine, HIV-associatednephropathy, transplant nephropathy, chronic urethral obstruction;hepatic diseases associated with excessive scarring and progressivesclerosis, including cirrhosis due to all etiologies, disorders of thebiliary tree, and hepatic dysfunction attributable to infections, suchas hepatitis virus or parasites; syndromes associated with pulmonaryfibrosis with consequential loss of gas exchange or ability toefficiently move air into and out of the lungs, including adultrespiratory distress syndrome, idiopathic pulmonary fibrosis, orpulmonary fibrosis due to infectious or toxic agents such as smoke,chemicals, allergens, or autoimmune disease; all collagen vasculardisorders of a chronic or persistent nature including progressivesystemic sclerosis, polymyositis, scleroderma, dermatomyositis,Raynaud's syndrome, or arthritic conditions such as rheumatoidarthritis; eye diseases associated with fibroproliferative states,including proliferative vitreoretinopathy of any etiology or fibrosisassociated with ocular surgery such as retinal reattachment, cataractextraction, or drainage procedures of any kind; excessive orhypertrophic scar formation in the dermis occurring during wound healingresulting from trauma or surgical wounds; disorders of thegastrointestinal tract associated with chronic inflammation, such asCrohn's disease or ulcerative colitis or adhesion formation as a resultof trauma or surgical wounds, polyposis or states post polyp surgery;chronic scarring of the peritoneum associated with endometriosis,ovarian disease, peritoneal dialysis, or surgical wounds; neurologicalconditions characterized by TGF-β production or enhanced sensitivity toTGF-β, including states post-traumatic or hypoxic injury, Alzheimer'sdisease, and Parkinson's disease; and diseases of the joints involvingscarring sufficient to impede mobility or produce pain, including statespost-mechanical or surgical trauma, osteoarthritis and rheumatoidarthritis.

The modulation of immune and inflammation systems by TGF-β includesstimulation of leukocyte recruitment, cytokine production, andlymphocyte effector function, and inhibition of T-cell subsetproliferation, β-cell proliferation, antibody formation, and monocyticrespiratory burst. Wahl et al., Immunol Today, 1989, 10, 258-61. TGF-βplays an important role in the pathogenesis of lung fibrosis which is amajor cause of suffering and death seen in pulmonary medicine based onits strong extracellular matrix inducing effect. The association ofTGF-β with human lung fibrotic disorders has been demonstrated inidiopathic pulmonary fibrosis, autoimmune lung diseases and bleomycininduced lung fibrosis. Nakao et al., J. Clin. Inv., 1999, 104, 5-11.

Other TGF-β disease states include inhibition of the intracellularsignaling pathway such as fibroproliferative diseases, including kidneydisorders associated with unregulated TGF-β activity and excessivefibrosis, including glomerulonephritis (GN), such as mesangialproliferative GN, immune GN, and crescentic GN. Other renal conditionsthat can be treated by inhibitors of TGF-β intracellular signalingpathway include diabetic nephropathy, renal interstitial fibrosis, renalfibrosis in transplant patients receiving cyclosporine, andHIV-associated nephropathy. Collagen vascular disorders that can betreated by inhibitors of TGF-β intracellular signaling pathway includeprogressive systemic sclerosis, polymyositis, sclerodema,dermnatomyositis, eosinophilic fascitis, morphea, or those associatedwith the occurrence of Raynaud's syndrome. Lung fibroses resulting fromexcessive TGF-β activity include adult respiratory distress syndrome,idiopathic pulmonary fibrosis, and interstitial pulmonary fibrosis oftenassociated with autoimmune disorders, such as systemic lupuserythematosus and scleroderma, chemical contact, or allergies. Anotherautoimmune disorder associated with fibroproliferative characteristicsis rheumatoid arthritis. Eye diseases associated with afibroproliferative condition include retinal reattachment surgeryaccompanying proliferative vitreoretinopathy, cataract extraction withintraocular lens implantation, and post glaucoma drainage surgery.

In addition to the disease states noted above, other diseases treatableor preventable by the administration of compounds or pharmaceuticallyacceptable salts thereof of the present invention include, but notlimited to, occlusive coronary thrombus formation resulting from eitherthrombolytic therapy or percutaneous transluminal coronary angioplasty,thrombus formation in the venous vasculature, disseminated intravascularcoagulopathy, a condition wherein there is rapid consumption ofcoagulation factors and systemic coagulation which results in theformation of life threatening thrombi occurring throughout themicrovasculature leading to widespread organ failure, hemmorhagicstroke, renal dialysis, blood oxygenation and cardiac catherization.

Compounds of the present invention can also be used in transcathetervascular therapies (TVT) including angioplasty, e.g., laser angioplastyand percutaneous transluminal coronary angioplasty (PTCA) proceduresemploying balloon catheters, and indwelling catheters; vascular graftingusing natural or synthetic materials, such as in saphenous vein coronarybypass grafts, dacron and venous grafts used for peripheral arterialreconstruction, etc.; placement of a mechanical shunt, such as a PTFEhemodialysis shunt used for arteriovenous communications; placement ofan intravascular stent, which may be metallic, plastic or abiodegradable polymer; or delivery of the compounds or pharmaceuticallyacceptable salt thereof to the lumen of a vessel via catheter, before,during or after angioplasty. See Schatz, U.S. Pat. No. 5,195,984;Palmaz, U.S. Pat. No. 4,739,762, the disclosures of which areincorporated herein by reference.

Compounds of the present invention can also be incorporated as a coatingon medical devices, such as stents, to prevent thrombus formation and tocontrol smooth muscle cell proliferation.

The stent or shunt useful in the method of the present invention cancomprise a biodegradable coating or porous non-biodegradable coating,having dispersed therein a sustained-release dosage form of the compoundor pharmaceutically acceptable salts thereof as a therapeutic agent. Inan alternative embodiment, a biodegradable stent or shunt may also havethe therapeutic agent impregnated in the stent or shunt matrix. Alsocontemplated is the use of a biodegradable stent or shunt with thetherapeutic agent impregnated therein which is further coated with abiodegradable coating or with a porous non-biodegradable coating havinga sustained release-dosage form dispersed therein. This embodiment ofthe invention provides a differential release rate of the therapeuticagent, i.e., there is a faster release of the therapeutic agent from thecoating followed by delayed release of the therapeutic agent that wasimpregnated in the stent or shunt matrix upon degradation of the stentor shunt matrix. The intravascular stent or shunt provides a mechanicalmeans of maintaining or providing an increase in luminal area of avessel, and the antiproliferative agent inhibits the vascular smoothmuscle cell proliferative response induced by the stent or shunt, whichcan cause occlusion of blood flow and coronary failure.

Compounds of the present invention can also be used to inhibit vascularsmooth muscle cell proliferation associated with procedural vasculartrauma due to organ transplantation, vascular surgery, angioplasty,shunt placement, stent placement or vascular grafting. This aspect ofthe invention generally comprises administering to a mammal, such as ahuman, that is subjected to the procedural trauma an effectiveantiproliferative amount of a compound of Formula I. Administration ofcompound of Formula I can be systemic, as by oral or parenteraladministration, or local, as to the site of the vascular trauma, orboth.

Compounds of the present invention can be used alone or in combinationwith other therapeutic or diagnostic agents. In certain preferredembodiments, the compounds of the invention are coadministered withother compounds typically prescribed for these conditions according togenerally accepted medical practice such as anticoagulant agents,thrombolytic agents, or other antithrombotics, including plateletaggregation inhibitors, tissue plasminogen activators, urokinase,prourokinase, streptokinase, heparin, aspirin, or warfarin. Thecompounds of the present invention may act in a synergistic fashion toprevent reocclusion following a successful thrombolytic therapy and/orreduce the time to reperfusion. In some instances, compounds of thepresent invention allow for reduced doses of the thrombolytic agents tobe used, and therefore minimize potential hemorrhagic side-effects.

In one particular embodiment, preferred compounds of Formula I are thosethat inhibit the phosphorylation of smad2 or smad3 by modulating TGF-βtype I or activin like kinase (ALK5) receptor. A particularly preferredgroup of compounds of Formula I are those having IC₅₀ of about 100 μM orless, with those having IC₅₀ of about 10 μM or less being moreparticularly preferred. Especially preferred compounds of Formula I arethose having IC₅₀ of about 1 μM or less.

Administration and Pharmaceutical Composition

Another aspect of the present invention provides a pharmaceuticalcomposition comprising at least one compound of the present inventiontogether with at least one pharmaceutically acceptable carrier (orexcipient), and optionally other therapeutic and/or prophylacticingredients.

Pharmaceutically acceptable carriers (or excipients) for therapeutic useare well known in the pharmaceutical art, and are described, forexample, in Remingtons Pharmaceutical Sciences, Mack Publishing Co. (A.R. Gennaro edit. 1985). For example, sterile saline andphosphate-buffered saline at physiological pH may be used.Preservatives, stabilizers, dyes and even flavoring agents may beprovided in the pharmaceutical composition. For example, sodiumbenzoate, sorbic acid and esters of p-hydroxybenzoic acid may be addedas preservatives. In addition, antioxidants and suspending agents may beused.

Compounds of the present invention can be administered in atherapeutically effective amount by any of the accepted modes ofadministration for agents that serve similar utilities. Thetherapeutically effective amount of the compound of the presentinvention can range broadly depending upon the desired affects and thetherapeutic indication. Therapeutically effective dosages can bedetermined by either in vitro or in vivo methods. For each particularcompound of the present invention, individual determinations can be madeto determine the optimal dosage required. The range of therapeuticallyeffective dosages is influenced by a variety of factors, such as theroute of administration, the therapeutic objectives, nature of thecondition, the severity of the condition, the particular subject to betreated, and the judgment of the practitioner.

The formulation will also depend on mode of administration. As thecompounds of the present invention are small molecules, they areconveniently administered orally by compounding them with suitablepharmaceutical excipients so as to provide tablets, capsules, syrups,and the like. Suitable formulations for oral administration can alsoinclude minor components such as buffers, flavoring agents and the like.Typically, the amount of active ingredient in the formulations will bein the range of 5%-95% of the total formulation, but wide variation ispermitted depending on the carrier. Suitable carriers include sucrose,pectin, magnesium stearate, lactose, peanut oil, olive oil, water, andthe like.

For injection by hypodermic needle, it may be assumed the dosage isdelivered into the body's fluids. For other routes of administration,the absorption efficiency is typically determined for each individualcompound by methods well known in pharmacology. Accordingly, it may benecessary for the therapist to titer the dosage and modify the route ofadministration as required to obtain the desired therapeutic effect. Thedetermination of effective dosage levels, that is, the dosage levelsnecessary to achieve the desired result, can be readily determined byone skilled in the art. Typically, applications of compound arecommenced at lower dosage levels, with dosage levels being increaseduntil the desired effect is achieved.

The compounds useful in the invention can also be administered throughsuppositories or other transmucosal vehicles. Typically, suchformulations include excipients that facilitate the passage of thecompound through the mucosa such as pharmaceutically acceptabledetergents. The compounds can also be administered topically, fortopical conditions, or in formulation intended to penetrate the skin.These include lotions, creams, ointments and the like which can beformulated by known methods in the art.

Compounds of the present invention can also be administered byinjection, including intravenous, intramuscular, subcutaneous orintraperitoneal injection. Typical formulations for such use are liquidformulations in isotonic vehicles such as Hank's solution or Ringer'ssolution. Alternative formulations include nasal sprays, liposomalformulations, slow-release formulations, and the like, as are known inthe art.

The dosages of the compounds of the invention depend on a number offactors which can vary from patient to patient. However, it is believedthat generally, the daily oral dosage will utilize 0.001-100 mg/kg totalbody weight, preferably from 0.01-50 mg/kg and more preferably about0.01 mg/kg-10 mg/kg. The dose regimen will likely vary, however,depending on the conditions being treated and the judgment of thepractitioner. It should be noted that the compounds of Formula I can beadministered as individual active ingredients, or as mixtures of severalembodiments of this formula. In addition, the inhibitors of TGF-β can beused as single therapeutic agents or in combination with othertherapeutic agents. Drugs that could be usefully combined with thesecompounds include natural or synthetic corticosteroids, particularlyprednisone and its derivatives, monoclonal antibodies targeting cells ofthe immune system, antibodies or soluble receptors or receptor fusionproteins targeting immune or non-immune cytokines, and small moleculeinhibitors of cell division, protein synthesis, or mRNA transcription ortranslation, or inhibitors of immune cell differentiation or activation.

The biological properties of the compounds of the present invention canbe readily characterized by methods that are well known in the art suchas, for example, by in vitro protease activity assays and in vivostudies to evaluate antithrombotic efficacy, and effects on hemostasisand hematological parameters, such as are illustrated in the examples.

Diagnostic applications of the compounds of the invention will typicallyutilize formulations in the form of solutions or suspensions. In themanagement of thrombotic disorders, the compounds of the invention canbe utilized in compositions such as tablets, capsules or elixirs fororal administration, suppositories, sterile solutions or suspensions orinjectable administration, and the like, or incorporated into shapedarticles. Subjects in need of treatment (typically mammalian) using thecompounds of the invention can be administered dosages that will provideoptimal efficacy. The dose and method of administration will vary fromsubject to subject and be dependent upon such factors as the type ofmammal being treated, its sex, weight, diet, concurrent medication,overall clinical condition, the particular compounds employed, thespecific use for which these compounds are employed, and other factorswhich those skilled in the medical arts will recognize.

Formulations of the compounds of this invention are prepared for storageor administration by mixing the compound having a desired degree ofpurity with physiologically acceptable carriers, excipients, stabilizersetc., and may be provided in sustained release or timed releaseformulations. Acceptable carriers or diluents for therapeutic use arewell known in the pharmaceutical field, and are described, for example,in Remington's Pharmaceutical Sciences, Mack Publishing Co., (A. R.Gennaro edit. 1985). Such materials are nontoxic to the recipients atthe dosages and concentrations employed, and include buffers such asphosphate, citrate, acetate and other organic acid salts, antioxidantssuch as ascorbic acid, low molecular weight (less than about tenresidues) peptides such as polyarginine, proteins, such as serumalbumin, gelatin, or immunoglobulins, hydrophilic polymers such aspolyvinylpyrrolidinone, amino acids such as glycine, glutamic acid,aspartic acid, or arginine, monosaccharides, disaccharides, and othercarbohydrates including cellulose or its derivatives, glucose, mannoseor dextrins, chelating agents such as EDTA, sugar alcohols such asmannitol or sorbitol, counterions such as sodium and/or nonionicsurfactants such as Tween, Pluronics or polyethyleneglycol. Thecompounds of the invention can be administered parenterally in aneffective amount within the dosage range of about 0.1 to 100 mg/kg,preferably about 0.5 to 50 mg/kg and more preferably about 1 to 20 mg/kgon a regimen in a single or 2 to 4 divided daily doses and/or continuousinfusion.

Typically, about 5 to 500 mg of a compound or mixture of compounds ofthis invention, as the free acid or base form or as a pharmaceuticallyacceptable salt, is compounded with a physiologically acceptablevehicle, carrier, excipient, binder, preservative, stabilizer, dye,flavor etc., as called for by accepted pharmaceutical practice. Theamount of active ingredient in these compositions is such that asuitable dosage in the range indicated is obtained.

Typical adjuvants which can be incorporated into tablets, capsules andthe like are binders such as acacia, corn starch or gelatin, andexcipients such as microcrystalline cellulose, disintegrating agentslike corn starch or alginic acid, lubricants such as magnesium stearate,sweetening agents such as sucrose or lactose, or flavoring agents. Whena dosage form is a capsule, in addition to the above materials it mayalso contain liquid carriers such as water, saline, or a fatty oil.Other materials of various types may be used as coatings or as modifiersof the physical form of the dosage unit. Sterile compositions forinjection can be formulated according to conventional pharmaceuticalpractice. For example, dissolution or suspension of the active compoundin a vehicle such as an oil or a synthetic fatty vehicle like ethyloleate, or into a liposome may be desired. Buffers, preservatives,antioxidants and the like can be incorporated according to acceptedpharmaceutical practice.

Dosage formulations of the compounds of this invention to be used fortherapeutic administration must be sterile. Sterility is readilyaccomplished by filtration through sterile membranes such as 0.2 micronmembranes, or by other conventional methods. Formulations typically willbe stored in lyophilized form or as an aqueous solution. The pH of thepreparations of this invention typically will be 3-11, more preferably5-9 and most preferably 7-8. It will be understood that use of certainof the foregoing excipients, carriers, or stabilizers will result in theformation of cyclic polypeptide salts. While the preferred route ofadministration is by injection, other methods of administration are alsoanticipated such as orally, intravenously (bolus and/or infusion),subcutaneously, intramuscularly, colonically, rectally, nasally,transdermally or intraperitoneally, employing a variety of dosage formssuch as suppositories, implanted pellets or small cylinders, aerosols,oral dosage formulations and topical formulations such as ointments,drops and dermal patches. The compounds of this invention are desirablyincorporated into shaped articles such as implants which may employinert materials such as biodegradable polymers or synthetic silicones,for example, Silastic, silicone rubber or other polymers commerciallyavailable.

Compounds of the invention can also be administered in the form ofliposome delivery systems, such as small unilamellar vesicles, largeunilamellar vesicles and multilamellar vesicles. Liposomes can be formedfrom a variety of lipids, such as cholesterol, stearylamine orphosphatidylcholines.

Compounds of this invention can also be delivered by the use ofantibodies, antibody fragments, growth factors, hormones, or othertargeting moieties, to which the compound molecules are coupled.Compounds of this invention can also be coupled with suitable polymersas targetable drug carriers. Such polymers can includepolyvinylpyrrolidinone, pyran copolymer,polyhydroxy-propyl-methacrylamide-phenol,polyhydroxyethyl-aspartamide-phenol, or polyethyleneoxide-polylysinesubstituted with palmitoyl residues. Furthermore, compounds of theinvention may be coupled to a class of biodegradable polymers useful inachieving controlled release of a drug, for example polylactic acid,polyglycolic acid, copolymers of polylactic and polyglycolic acid,polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters,polyacetals, polydihydropyrans, polycyanoacrylates and cross linked oramphipathic block copolymers of hydrogels. Polymers and semipermeablepolymer matrices may be formed into shaped articles, such as valves,stents, tubing, prostheses and the like.

EXAMPLES

Additional objects, advantages, and novel features of this inventionwill become apparent to those skilled in the art upon examination of thefollowing examples thereof. It should be appreciated that these examplesare not to be considered as limiting the scope of the invention, butmerely as being illustrative and representative thereof.

Example 1 Preparation of[3-fluoro-2-(2-fluoro-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine

Step A: Preparation of 3-diazoquinoline

3-Aminoquinoline (10 g, 69 mmol) was dissolved in 70 mL of conc. HCl ina 250 mL round bottomed flask and cooled to 0° C. Sodium nitrite (9.6 g,139 mmol) in 20 mL water was added drop wise over 0.5 h whilemaintaining the reaction temperature between 0-5° C. After stirring foranother 0.5 h, fluoroboric acid (20 mL) was added and the stirringcontinued for another 0.5 h. The solid was filtered and dried undervacuum to yield the title compound (15.2 g, 90%).

Step B: Preparation of 3-fluoroquinoline

The tetrafluoroborate salt from Step A was suspended in 100 mL oftoluenein 250 mL round bottomed flask equipped with a magnetic stirrer andheated to 110° C. An exothermic reaction occurred and the flask wasmaintained at that temperature until the evolution of gas stopped. Thesolvent was removed and the residue was basified to pH 7 using potassiumcarbonate. The resulting solution was extracted with 3×50 mL of ethylacetate, and the organic layers were combined, dried over magnesiumsulfate and concentrated in a rotary evaporator. The crude compound waspurified by silica gel column chromatography using 9:1 hexane:ethylacetate to yield the title compound (6.3 g, 68%). MS (ES) 148.1 (M+H)⁺.

Step C: Preparation of 3-fluoro-4-iodo-quinoline

To a 0° C. solution of diisopropyl amine (2.8 mL, 27 mmol) intetrahydrofuran (80 mL) was added dropwise butyl lithium (8.1 mL, 2.5 Msolution in hexane, 27 mmol). After 10 min., the flask was cooled to−78° C. and 3-fluoro quinoline from Step B (3 g, 20 mmol) in 20 mL oftetrahydrofuran was added over 15 min. period. The stirring wascontinued for an additional 4 h., after which iodine (5.4 g, 21 mmol) in10 mL of THF (tetrahydrofuran) was added and the reaction temperaturewas maintained at −78° C. for additional 2 h. The reaction was quenchedby adding a mixture of water and tetrahydrofuran (25 mL, 1:9), and theresulting solution was extracted with ethyl acetate. The organic layerswere combined, washed with water and brine. Purification by silica gelcolumn gave the title compound (2.26 g, 40%). MS (ES) 148.1 (M+H)⁺.

Step D: Preparation of 4-chloro-3-fluoro-2-iodo-quinoline

To a 0° C. solution of diisopropyl amine (0.215 mL, 2.1 mmol) in THF (5mL) was added drop wise a solution of butyl lithium (0.615 mL, 2.5 Msolution in hexane, 2.1 mmol). After 10 min. the flask was cooled to−78° C. and a solution of 3-fluoro-4-iodo-quinoline (from Step C, 0.412g, 1.5 mmol) in 2 mL of tetrahydrofuran was added over 5 min. Thestirring was continued for an additional 2 h., after which a solution ofhexachloroethane (0.356 g, 1.5 mmol) in 1 mL of tetrahydrofuran wasadded and the reaction mixture was stirred at −78° C. for additional 2h. The reaction mixture was diluted with a mixture of water andtetrahydrofuran (10 mL, 1:9) and extracted with ethyl acetate. Theorganic layers were combined, washed with water and brine, and purifiedby silica gel column chromatography to yield the title compound (0.318g, 70%). MS (ES) 308.1 (M+H)⁺.

Step E: Preparation of 4-chloro-3-fluoro-2-(2-fluoro-phenyl)-quinoline

To a solution of 4-chloro-3-fluoro-2-iodo-quinoline (from step D, 0.3 g,0.9 mmol) in toluene (5 mL) was added n-butanol (2 mL), water (3 mL) andcesium carbonate (1.14 g, 3.5 mmol). The resulting mixture was degassedfor 5 min. and tetrakis(triphenyl-phosphine)palladium(0) (5 mg, 5 mol%)and 2-fluorophenyl boronic acid (0.286 g, 2 mmol) were added. Thereaction mixture was heated to 50° C. for 24 h., after which it wasextracted with ethyl acetate. The organic layer was dried andconcentrated in a rotary evaporator. The residue was purified by silicagel column chromatography using ethyl acetate/hexane gradient to yieldthe title compound (0.160 g, 48%). MS (ES) 336.1 (M+H)⁺.

Step F: Preparation of 3-fluoro-2-(2-fluoro-phenyl)-4-iodo-quinoline

To a solution of 4-chloro-3-fluoro-2-(2-fluoro-phenyl)-quinoline (fromstep E, 0.160 g, 0.47 mmol) in tetrahydrofuran (3 mL) at −78° C. wasadded butyl lithium (0.6 mL, 2.5 M in hexane, 1.3 mmol). The mixture wasstirred for 3 h. and iodine (0.148 g, 0.58 mmol) in 0.8 mL of THF wasadded. The reaction mixture was stirred for an additional 2 h. Thereaction mixture was diluted with water (20 mL) and extracted with ethylacetate (2×25 mL). The organic layers were combined, dried over sodiumsulfate, concentrated and purified by silica gel column chromatographyto yield the title compound (0.148 g, 84%). MS (ES) 368.1 (M+H)⁺.

Step G: Preparation of[3-fluoro-2-(2-fluoro-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine

To a solution of 3-fluoro-2-(2-fluoro-phenyl)-4-iodo-quinoline (fromStep F, 0.02 g, 0.05 mmol) in toluene (1 mL) was added S-BINAP (0.007 g,0.01 mmol), sodium tert-butoxide (0.105 g, 1 mmol) and 4-aminopyridine(0.015 g, 0.05 mmol). The resulting mixture was degassed for 5 min.,after which tris(dibenzylidineacetone) dipalladium(0) (0.0025 g, 0.0025mmol) was added. The reaction mixture was heated to 110° C. for 48 h.and then diluted with water and extracted with ethyl acetate. Theorganic layer was dried over sodium sulfate and concentrated in rotaryevaporator. The residue was purified using HPLC over a gradient ofacetonitrile and water containing 0.1 % trifluoroacetic acid to yieldthe desired compound (0.012 g, 65%). MS (ES) 334.1 (M+H)⁺.

Example 2 Preparation of[3-Fluoro-2-(2-fluoro-phenyl)-guinolin-4-yl]-pyridin-3-yl-amine

To a solution of 3-fluoro-2-(2-fluoro-phenyl)-4-iodo-quinoline (fromExample 1, Step F, 0.02 g, 0.05 mmol) in dioxane (1 mL) was added cesiumcarbonate (0.05 g, 0.15 mmol) and Xanphos (0.0045 g, 0.001 mmol). Theresulting mixture was degassed for 5 min andtris(dibenzylidineacetone)dipalladium(0) (0.0025 g, 0.0025 mmol) wasadded. The reaction mixture was stirred at 100° C. for 3 h, after whichit was diluted with water and extracted with ethyl acetate. The organiclayer was dried over sodium sulfate and concentrated in rotaryevaporator. The residue was purified using HPLC over a gradient ofacetonitrile and water containing 0.1 % trifluoroacetic acid to yieldthe desired compound (0.012 g, 65%). MS (ES) 334.1 (M+H)⁺.

Example 3 Preparation of(2-benzo[1,3]dioxol-5-yl-3-fluoro-quinolin-4-yl)-pyridin-4-yl-amine

Step A: Preparation of2-benzo[1,3]dioxol-5-yl-4-chloro-3-fluoro-quinoline

To a solution of 4-chloro-3-fluoro-2-iodo-quinoline (from Example 1,step D, 0.127 g, 0.41 mmol) in toluene (5 mL) was added n-butanol (2mL), water (3 mL) and cesium carbonate (0.404 g, 1.2 mmol). Theresulting mixture was degassed for 5 min., after whichtetrakis(triphenylphosphine)palladium(0) (0.002 g, 5 mol%) andbenzo[1,3]dioxole-5-boronic acid (0.086 g, 0.4 mmol) were added. Thereaction mixture was stirred at 50° C. for 24 h. and then extracted withethyl acetate. The organic layer was dried and concentrated in a rotaryevaporator. The residue was purified by silica gel column chromatographyusing ethyl acetate/hexane gradient to yield the title compound (0.070g, 57%). MS (ES) 302 (M+H)⁺.

Step B: Preparation of 2-benzo[1,3]dioxol-5-yl-3-fluoro-4-iodo-quinoline

To a −78° C. solution of2-benzo[1,3]dioxol-5-yl-4-chloro-3-fluoro-quinoline (from step A, 0.073g, 0.24 mmol) in tetrahydrofuran (2 mL) was added butyl lithium (0.36mL, 2.5 M in hexane, 0.6 mmol). The resulting reaction mixture wasstirred for 3 h., after which a solution of iodine (0.06 g, 0.24 mmol)in 0.8 mL of THF was added. The reaction mixture was stirred for anadditional 2 h. and diluted with water (20 mL). The aqueous layer wasextracted with ethyl acetate (2×25 mL). The combined organic layers weredried over sodium sulfate, concentrated and purified via silica gelcolumn chromatography to give the title compound (0.046 g, 48%). MS (ES)393.8 (M+H)⁺.

Step C: Preparation of(2-benzo[1,3]dioxol-5-yl-3-fluoro-quinolin-4-yl)-pyridin-4-yl-amine

To a solution of 2-benzo[1,3]dioxol-5-yl-3-fluoro-4-iodo-quinoline (fromStep B, 0.04 g, 0.1 mmol) in dioxane (1 mL) was added cesium carbonate(0.1 g, 0.3 mmol) and Xanphos (0.0045 g, 0.001 mmol). The mixture wasdegassed for 5 min., and then tris(dibenzylidineacetone)dipalladium(0)(0.0025 g, 0.0025 mmol) was added. The reaction mixture was stirred at100° C. for 2 h., diluted with water and extracted with ethyl acetate.The organic layer was dried over sodium sulfate and concentrated inrotary evaporator. The residue was purified via HPLC over a reversephase column using a gradient of acetonitrile and water containing 0.1%trifluoroacetic acid to yield the desired compound (0.009 g, 24%). MS(ES) 360.0 (M+H)⁺.

Assay Methods

The following assay methods were used to evaluate the compounds of thepresent invention.

The autophosphorylation of GST-ALK5 was developed for primary screeningof the compounds that inhibit TGF-β signaling by interacting with ALK5.HA-ALK5 assay is a secondary screening assay to confirm the inhibitorycompounds that were selected from the primary screening, and also forthe determination of IC₅₀ for each compound.

Autophosphorylation Assay of GST-ALK5

The cytoplasmic domain of ALK5 was fused to glutathione S-transferase(GST) and the GST-ALK5 fusion protein was expressed in a baculovirusexpression system. GST-ALK5 was isolated with glutathione Sepharose 4Bbeads (Pharmacia Biotech, Sweden) and stored at −80° C. until use.

For detection of GST-ALK5 autophosphorylation and screening ofinhibitory compounds, an aliquot of GST-ALK5 in 1× kinase bufferincluding [³³P]-γ-ATP was added to 96-well plates in the presence orabsence of compounds. The mixture was then incubated at room temperaturefor 30 min and transferred to each well of a Filterplate with vacuum.The Filterplate was then washed 3 times and radioactivity in each wellwas counted in a Packard TopCount.

In Vitro Kinase of HA-ALK5

An expression construct containing full-length ALK5 C-terminally taggedwith HA was transfected into COS7 cells, and HA-ALK5 was isolated byimmunoprecipitation with anti-HA antibodies. Aliquots ofimmunoprecipitated HA-ALK5 in 1× kinase buffer plus [³³P]-γ-ATP wasadded to 96-well plates in the presence or absence of differentconcentrations of testing compounds, and incubated at room temperaturefor 60 min. The reaction mixture was then transferred to a Filterplate.The plate was washed three times and radioactivity in each well counted.IC₅₀ for each compound was determined using the Prism3 program.

ELISA Assay for TGF-β Stimulated Smad2 Phosphorylation

Serum-starved normal human lung fibroblasts (NHLF) in 24-well plate weretreated with or without different concentrations of testing compoundsfor 30 min. The cells were then stimulated with TGF-β for one hour.After fixing, permeabilizing, and blocking, the cells were incubatedwith phospho-Smad2 specific antibodies followed by HRP-conjugatedsecondary antibody. The extent of Smad2 phosphorylation was thendetected using HRP substrate and read with an ELISA plate reader. IC₅₀for each testing compound was determined using the PRISM3 program.

ELISA Assay for TGF-β Stimulated PAI-1 Secretion

Serum-starved NHLF in 24-well plates were treated with or withoutdifferent concentrations of testing compounds for 30 min. The cells werethen stimulated with TGF-β and incubated in a 37° C. incubator for 24hours. The media were collected and added to 96-well plates coated withanti-PAI-1 antibodies. The secreted PAI-1 was then detected with anotherPAI-1 specific antibody followed by HRP-conjugated secondary antibody.The secretion of PAI-1 was detected using HRP substrate and read with anELISA plate reader. IC₅₀ for each testing compound was determined usingthe PRISM3 program.

SIRCOL Collagen Assay for TGF-β Stimulated Cells

Serum-starved NHLF in 24-well plates were treated with or withoutdifferent concentrations of testing compounds for 30 min. The cells werethen stimulated with TGF-β and incubated for 24 hours. The media werecollected and SIRCOL dye reagent was added. After spinning and washing,the pellets were resuspended in alkali reagent and read with an ELISAplate reader. IC₅₀ for each testing compound was determined using thePRISM3 program.

Detection of TGF-β Stimulated Fibronectin Expression

Serum-starved NHLF in 24-well plates were treated with or withoutdifferent concentrations of testing compounds for 30 min. The cells werethen stimulated with TGF-β and incubated for 24 hours. After washing andfixing, the secreted fibronectin was incubated with fibronectin specificantibodies, followed by incubation with biotin-labeled secondaryantibody, streptavidin-peroxidase and HRP substrate. The signal was thendetected using an ELISA reader. IC₅₀ for each testing compound wasdetermined using the PRISM3 program.

Results

Some of the representative compounds having IC₅₀ value of <10 μM forTGF-β (ALK5) phosphorylation and in cell-based assays include compoundNos. 7, 51, 55, and 62-66 in Table 1. In addition, these compoundsexhibited >50-100-fold selectivity for ALK5 (TGF-β) vs. closely relatedALK6 and p38 kinases.

P-Smad2, PAI-1, collagen and fibronectin assays are cell-based assaysthat are used for determination of functional activities of thecompounds from the secondary screening. Since all of the molecules aretargets of TGF-β signaling, the data demonstrated that the compoundsspecifically inhibit TGF-β mediated signal transduction.

Without further description, it is believed that one of ordinary skillin the art can, using the preceding description and the illustrativeexamples, make and utilize the compounds of the present invention andpractice the claimed methods. It should be understood that the foregoingdiscussion and examples merely present a detailed description of certainpreferred embodiments. It will be apparent to those of ordinary skill inthe art that various modifications and equivalents can be made withoutdeparting from the spirit and scope of the invention. All the patents,journal articles and other documents discussed or cited above are hereinincorporated by reference.

1. A compound of the formula:

or a pharmaceutically acceptable salt or a prodrug thereof, wherein Pgis hydrogen, alkyl or a nitrogen protecting group; n is an integer from0 to 3; each R¹ substituent is independently selected from the groupconsisting of —R², -T-R², and -V-T-R; each R² is independently selectedfrom the group consisting of C₁₋₃ aliphatic, hydroxy, —N(R³)₂, halo,cyano, —OR⁴, —C(O)R⁴, —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴, —N(R³)C(O)R⁴,—N(R³)CO₂R⁴, —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂, —N(R³)C(O)N(R³)₂,—OC(O)R⁴, phenyl which is optionally substituted with 1-3 R⁵, 5-6membered heterocyclyl which is optionally substituted with 1-3 R⁵, and5-6 membered heteroaryl which is optionally substituted with 1-3 R⁵;each T is independently C₁₋₅ alkylidene that is optionally interruptedby —O—, —C(O)—, —S—, —S(O)—, —S(O)₂—, or —N(R³)—; each V isindependently selected from the group consisting of —O—, —S—, —S(O)—,—S(O)₂—, —C(O)—, —N(R³)—, —N(R³)C(O)—, or —N(R³)C(O)₂—, —N(R³)S(O)₂—,—C(O)N(R³)—, —S(O)₂N(R³)—, —N(R³)C(O)N(R³)—, and —OC(O)—; each R³ isindependently selected from the group consisting of hydrogen, C₁₋₆aliphatic, —C(O)R⁴, —C(O)₂R⁴, and —SO₂R⁴, or two R³ on the same nitrogentogether with their intervening nitrogen form a 5-6 memberedheterocyclyl or heteroaryl ring having 1-3 ring heteroatoms selectedfrom nitrogen, oxygen or sulfur; each R⁴ is independently selected fromthe group consisting of C₁₋₆ aliphatic, phenyl or a 5-6 memberedheteroaryl or heterocyclyl having 1-3 ring heteroatoms selected fromnitrogen, oxygen or sulfur; R⁸ is selected from the group consisting of:(a) aryl, heteroaryl, and cycloalkyl, each of which is optionallysubstituted with 1-3 R⁵, and (b) alkyl; each R⁵ is independentlyselected from the group consisting of C₁₋₆ aliphatic, halo, —OH,—N(R³)₂, cyano, —OR⁴, —C(O)R⁴, —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴,—N(R³)C(O)R⁴, —N(R³)CO₂R⁴, —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂,—N(R³)C(O)N(R³)₂, —OC(O)R⁴, —OC(O)N(R³)₂, phenyl, 5-6 memberedheterocyclyl and 5-6 membered heteroaryl, or two adjacent R⁵ on an aryl,cycloalkyl, or heteroaryl ring are taken together with their interveningatoms to form a 5-6 membered fused ring having 0-2 heteroatoms selectedfrom nitrogen, oxygen or sulfur; each R⁶ is independently selected fromthe group consisting of hydrogen, halo, C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄monoalkylamino and C₁₋₄ dialkylamino; and R⁷ is selected from the groupconsisting of hydrogen, halo, —OH, —N(R³)₂, cyano, —OR⁴, —C(O)R⁴,—CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴, —N(R³)C(O)R⁴, —N(R³)CO₂R⁴, —N(R³)SO₂R⁴,—C(O)N(R³)₂, —SO₂N(R³)₂, —N(R³)C(O)N(R³)₂, and —OC(O)R⁴.
 2. The compoundaccording to claim 1, wherein Pg is hydrogen.
 3. The compound accordingto claim 2, wherein R⁸ is selected from the group consisting of phenyl,naphthyl, pyridyl, thienyl, furyl, each of which is optionallysubstituted with 1-3 R⁵, cyclohexyl, cyclopentyl, cyclopropyl, andt-butyl.
 4. The compound according to claim 3, wherein R⁸ is selectedfrom the group consisting of 2-fluorophenyl, benzo[1,3]dioxol-5-yl,2-trifluoromethylphenyl, 2-chlorophenyl, pyridin-4-yl, 2-methylphenyl,2-methoxyphenyl, 3-fluorophenyl, 3-chlorophenyl, 5-chloro-thiophen-2-yl,5-chloro-furan-2-yl, 5-methyl-thiophen-2-yl, 5-methyl-furan-2-yl,4-methyl-thiophen-2-yl, 2,3-dihydro-benzofuran-5-yl,2-methylsulfanylphenyl, 4-fluorophenyl, 2-methanesulfinylphenyl,4-methoxyphenyl, 2-cyanophenyl, 2-amidophenyl, 4-hydroxyphenyl,3-aminophenyl, 3-methoxyphenyl, 4-cyanophenyl, 2,6-dichlorophenyl,phenyl, 4-chlorophenyl, cyclohexyl, cyclopropyl, cyclopentyl,4-methylsulfanylphenyl, t-butyl, 4-amidophenyl, naphthalen-2-yl,4-methanesulfinylphenyl, and 2-bromophenyl.
 5. The compound according toclaim 4, wherein R⁶ is hydrogen.
 6. The compound according to claim 5,wherein R⁷ is hydrogen.
 7. The compound according to claim 6, whereineach R¹ is independently selected from the group consisting of: (a)thienyl which is optionally substituted with 1-3 R⁵; (b) furyl which isoptionally substituted with 1-3 R⁵; (c) pyrrolyl which is optionallysubstituted with 1-3 R⁵; (d) phenyl which is optionally substituted with1-3 R⁵; (e) halo, (f) —OR⁴, (g) —N(R³)₂; and (h) —N(R³)-T-OR⁴.
 8. Thecompound according to claim 7 of the formula:


9. The compound according to claim 8, wherein R¹ is selected from thegroup consisting of furyl, pyrrolyl, thienyl, phenyl which is optionallysubstituted with R⁵, and —N(R³)-T-OR⁴.
 10. The compound according toclaim 7 of the formula:


11. The compound according to claim 10, wherein R¹ is selected from thegroup consisting of halo, —OR⁴, —N(R³)₂, and —N(R³)-T-OR⁴.
 12. Thecompound according to claim 1, wherein R⁸ is selected from the groupconsisting of phenyl, naphthyl, pyridyl, thienyl, furyl, each of whichis optionally substituted with 1-3 R⁵, cyclohexyl, cyclopentyl,cyclopropyl, and t-butyl.
 13. The compound according to claim 12 of theformula:


14. The compound according to claim 12, wherein Pg, R⁶ and R⁷ arehydrogen.
 15. The compound according to claim 14, wherein each R¹ isindependently selected from the group consisting of: (a) thienyl whichis optionally substituted with 1-3 R⁵; (b) furyl which is optionallysubstituted with 1-3 R⁵; (c) pyrrolyl which is optionally substitutedwith 1-3 R⁵; (d) phenyl which is optionally substituted with 1-3 R⁵; (e)halo, (f) —OR⁴, (g) —N(R³)₂; and (h) —N(R³)-T-OR⁴.
 16. The compoundaccording to claim 15, wherein n is
 1. 17. The compound according toclaim 1 of the formula:


18. The compound according to claim 1 of the formula:


19. The compound according to claim 1, wherein said compound is selectedfrom the group consisting of:[3-Fluoro-2-(2-methylsulfanyl-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine,[3-Fluoro-2-(2-methanesulfinyl-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine,2-[3-Fluoro-4-(pyridin-4-ylamino)-quinolin-2-yl]-benzonitrile,2-[3-Fluoro-4-(pyridin-4-ylamino)-quinolin-2-yl]-benzamide,[3-Fluoro-2-(4-fluoro-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine,[3-Fluoro-2-(4-methoxy-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine,[2-(3-Chloro-phenyl)-3-fluoro-quinolin-4-yl]-pyridin-4-yl-amine,[3-Fluoro-2-(3-fluoro-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine,4-[3-Fluoro-4-(pyridin-4-ylamino)-quinolin-2-yl]-phenol,[3-Fluoro-2-(3-methoxy-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine,[2-(2,6-Dichloro-phenyl)-3-fluoro-quinolin-4-yl]-pyridin-4-yl-amine,[2-(4-Chloro-phenyl)-3-fluoro-quinolin-4-yl]-pyridin-4-yl-amine,[2-(3-Amino-phenyl)-3-fluoro-quinolin-4-yl]-pyridin-4-yl-amine,4-[3-Fluoro-4-(pyridin-4-ylamino)-quinolin-2-yl]-benzonitrile,(3-Fluoro-2-phenyl-quinolin-4-yl)-pyridin-4-yl-amine,(2-Cyclohexyl-3-fluoro-quinolin-4-yl)-pyridin-4-yl-amine,(2-Cyclopropyl-3-fluoro-quinolin-4-yl)-pyridin-4-yl-amine,(2-tert-Butyl-3-fluoro-quinolin-4-yl)-pyridin-4-yl-amine,(3-Fluoro-2-naphthalen-2-yl-quinolin-4-yl)-pyridin-4-yl-amine,(2-Cyclopentyl-3-fluoro-quinolin-4-yl)-pyridin-4-yl-amine,[3-Fluoro-2-(4-methylsulfanyl-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine,4-[3-Fluoro-4-(pyridin-4-ylamino)-quinolin-2-yl]-benzamide,(3-Fluoro-2-phenyl-quinolin-4-yl)-pyridin-4-yl-amine,[3-Fluoro-2-(4-methanesulfinyl-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine,[3-Fluoro-2-(2-fluoro-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine,(2-Benzo[1,3]dioxol-5-yl-3-fluoro-quinolin-4-yl)-pyridin-4-yl-amine,(3-Fluoro-2-pyridin-4-yl-quinolin-4-yl)-pyridin-4-yl-amine,(3-Fluoro-2-o-tolyl-quinolin-4-yl)-pyridin-4-yl-amine,[2-(2-Bromo-phenyl)-3-fluoro-quinolin-4-yl]-pyridin-4-yl-amine,[3-Fluoro-2-(2-trifluoromethyl-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine,[2-(2-Chloro-phenyl)-3-fluoro-quinolin-4-yl]-pyridin-4-yl-amine,[3-Fluoro-2-(2-methoxy-phenyl)-quinolin-4-yl]-pyridin-4-yl-amine,3-Fluoro-2-(2-fluoro-phenyl)-N6-methyl-N4-pyridin-4-yl-quinoline-4,6-diamine,3-Fluoro-2-(2-fluoro-phenyl)-N6-(2-methoxy-ethyl)-N4-pyridin-4-yl-quinoline-4,6-diamine,[3-Fluoro-2-(2-fluoro-phenyl)-6-thiophen-3-yl-quinolin-4-yl]-pyridin-4-yl-amine,2-Benzo[1,3]dioxol-5-yl-3-fluoro-N6-methyl-N4-pyridin-4-yl-quinoline-4,6-diamine,2-Benzo[1,3]dioxol-5-yl-3-fluoro-N6-(2-methoxy-ethyl)-N4-pyridin-4-yl-quinoline-4,6-diamine,[3-Fluoro-2-(2-fluoro-phenyl)-6-furan-2-yl-quinolin-4-yl]-pyridin-4-yl-amine,(2-Benzo[1,3]dioxol-5-yl-7-bromo-3-fluoro-quinolin-4-yl)-pyridin-4-yl-amine,2-Benzo[1,3]dioxol-5-yl-3-fluoro-N7-methyl-N4-pyridin-4-yl-quinoline-4,7-diamine,2-Benzo[1,3]dioxol-5-yl-3-fluoro-N7-(2-methoxy-ethyl)-N4-pyridin-4-yl-quinoline-4,7-diamine,(2-Benzo[1,3]dioxol-5-yl-3-fluoro-7-methoxy-quinolin-4-yl)-pyridin-4-yl-amine,[3-Fluoro-2-(2-fluoro-phenyl)-7-methoxy-quinolin-4-yl]-pyridin-4-yl-amine,3-Fluoro-2-(2-fluoro-phenyl)-N7-methyl-N4-pyridin-4-yl-quinoline-4,7-diamine,[2-(3-Bromo-phenyl)-3-fluoro-quinolin-4-yl]-pyridin-4-yl-amine,(2-Chloro-pyridin-4-yl)-[3-fluoro-2-(5-methyl-furan-2-yl)-quinolin-4-yl]-amine,(2-Chloro-pyridin-4-yl)-[2-(5-chloro-thiophen-2-yl)-3-fluoro-quinolin-4-yl]-amine,(2-Chloro-pyridin-4-yl)-[3-fluoro-2-(3-fluoro-phenyl)-quinolin-4-yl]-amine,[3-Fluoro-2-(5-methyl-furan-2-yl)-quinolin-4-yl]-methyl-pyridin-4-yl-amine,and(2-Chloro-pyridin-4-yl)-[2-(5-chloro-thiophen-2-yl)-3-fluoro-quinolin-4-yl]-methyl-amine.20. A method of treating a TGF-β mediated disorder in a patient in needthereof, comprising administering to said patient a compound of theformula:

or a pharmaceutically acceptable salt or a prodrug thereof, wherein Pgis hydrogen, alkyl or a nitrogen protecting group; n is an integer from0 to 3; each R¹ substituent is independently selected from the groupconsisting of —R², -T-R², and -V-T-R; each R² is independently selectedfrom the group consisting of C₁₋₃ aliphatic, hydroxy, —N(R³)₂, halo,cyano, —OR⁴, —C(O)R⁴, —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴, —N(R³)C(O)R⁴,—N(R³)CO₂R⁴, —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂, —N(R³)C(O)N(R³)₂,—OC(O)R⁴, phenyl which is optionally substituted with 1-3 R⁵, 5-6membered heterocyclyl which is optionally substituted with 1-3 R⁵, and5-6 membered heteroaryl which is optionally substituted with 1-3 R⁵;each T is independently C₁₋₅ alkylidene that is optionally interruptedby —O—, —C(O)—, —S—, —S(O)—, —S(O)₂—, or —N(R³)—; each V isindependently selected from the group consisting of —O—, —S—, —S(O)—,—S(O)₂—, —C(O)—, —N(R³)—, —N(R³)C(O)—, or —N(R³)C(O)₂—, —N(R³)S(O)₂—,—C(O)N(R³)—, —S(O)₂N(R³)—, —N(R³)C(O)N(R³)—, and —OC(O)—; each R³ isindependently selected from the group consisting of hydrogen, C₁₋₆aliphatic, —C(O)R⁴, —C(O)₂R⁴, and —SO₂R⁴, or two R³ on the same nitrogentogether with their intervening nitrogen form a 5-6 memberedheterocyclyl or heteroaryl ring having 1-3 ring heteroatoms selectedfrom nitrogen, oxygen or sulfur; each R⁴ is independently selected fromthe group consisting of C₁₋₆ aliphatic, phenyl or a 5-6 memberedheteroaryl or heterocyclyl having 1-3 ring heteroatoms selected fromnitrogen, oxygen or sulfur; R⁸ is selected from the group consisting of:(a) aryl, heteroaryl, and cycloalkyl, each of which is optionallysubstituted with 1-3 R⁵, and (b) alkyl; each R⁵ is independentlyselected from the group consisting of C₁₋₆ aliphatic, halo, —OH,—N(R³)₂, cyano, —OR⁴, —C(O)R⁴, —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴,—N(R³)C(O)R⁴, —N(R³)CO₂R⁴, —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂,—N(R³)C(O)N(R³)₂, —OC(O)R⁴, —OC(O)N(R³)₂, phenyl, 5-6 memberedheterocyclyl and 5-6 membered heteroaryl, or two adjacent R⁵ on an aryl,cycloalkyl, or heteroaryl ring are taken together with their interveningatoms to form a 5-6 membered fused ring having 0-2 heteroatoms selectedfrom nitrogen, oxygen or sulfur; each R⁶ is independently selected fromthe group consisting of hydrogen, halo, C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄monoalkylamino and C₁₋₄ dialkylamino; and R⁷ is selected from the groupconsisting of hydrogen, halo, —OH, —N(R³)₂, cyano, —OR⁴, —C(O)R⁴,—CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴, —N(R³)C(O)R⁴, —N(R³)CO₂R⁴, —N(R³)SO₂R⁴,—C(O)N(R³)₂, —SO₂N(R³)₂, —N(R³)C(O)N(R³)₂, and —OC(O)R⁴.
 21. The methodaccording to claim 20, wherein the disorder is a cardiovascular diseaseselected from the group consisting of atherosclerosis, thrombosis,myocardial infarction, stroke, congestive heart failure, dilatedcardiomyopathy, vascular stenosis associated with atherosclerosis,angioplasty treatment, surgical incisions and mechanical trauma.
 22. Themethod according to claim 20, wherein the disorder is fibroproliferativediseases selected from the group consisting of: kidney diseasesassociated with fibrosis and/or sclerosis, glomerulonephritis of alletiologies, diabetic nephropathy, and renal interstitial fibrosis,hypertension, complications of drug exposure, HIV-associatednephropathy, transplant nephropathy, chronic urethral obstruction;hepatic diseases associated with excessive scarring and progressivesclerosis, cirrhosis due to all etiologies, disorders of the biliarytree, and hepatic dysfunction attributable to infections such ashepatitis virus or parasites; syndromes associated with pulmonaryfibrosis, adult respiratory distress syndrome, idiopathic pulmonaryfibrosis, pulmonary fibrosis; collagen vascular disorders; Raynaud'ssyndrome, or arthritic conditions; eye diseases associated withfibroproliferative states, disorders of the gastrointestinal tractassociated with chronic inflammation; Crohn's disease, and chronicscarring of the peritoneum associated with endometriosis, ovariandisease, peritoneal dialysis or surgical wounds.
 23. A pharmaceuticalcomposition comprising: (a) a pharmaceutically acceptable carrier; and(b) a compound of the formula:

or a pharmaceutically acceptable salt or a prodrug thereof, wherein Pgis hydrogen, alkyl, or a nitrogen protecting group; n is an integer from0 to 3; each R¹ substituent is independently selected from the groupconsisting of —R², -T-R², and -V-T-R; wherein each R² is independentlyselected from the group consisting of C₁₋₃ aliphatic, hydroxy, —N(R³)₂,halo, cyano, —OR⁴, —C(O)R⁴, —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴,—N(R³)C(O)R⁴, —N(R³)CO₂R⁴, —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂,—N(R³)C(O)N(R³)₂, —OC(O)R⁴, phenyl which is optionally substituted with1-3 R⁵, 5-6 membered heterocyclyl which is optionally substituted with1-3 R⁵, and 5-6 membered heteroaryl which is optionally substituted with1-3 R⁵; each T is independently C₁₋₅ alkylidene that is optionallyinterrupted by —O—, —C(O)—, —S—, —S(O)—, —S(O)₂—, or —N(R³)—; each V isindependently selected from the group consisting of —O—, —S—, —S(O)—,—S(O)₂—, —C(O)—, —N(R³)—, —N(R³)C(O)—, or —N(R³)C(O)₂—, —N(R³)S(O)₂—,—C(O)N(R³)—, —S(O)₂N(R³)—, —N(R³)C(O)N(R³)—, and —OC(O)—; each R³ isindependently selected from the group consisting of hydrogen, C₁₋₆aliphatic, —C(O)R⁴, —C(O)₂R⁴, and —SO₂R⁴, or two R³ on the same nitrogentogether with their intervening nitrogen form a 5-6 memberedheterocyclyl or heteroaryl ring having 1-3 ring heteroatoms selectedfrom nitrogen, oxygen or sulfur; each R⁴ is independently selected fromthe group consisting of C₁₋₆ aliphatic, phenyl or a 5-6 memberedheteroaryl or heterocyclyl having 1-3 ring heteroatoms selected fromnitrogen, oxygen or sulfur; R⁸ is selected from the group consisting of:(a) aryl, heteroaryl, and cycloalkyl, each of which is optionallysubstituted with 1-3 R⁵, and (b) alkyl; wherein each R⁵ is independentlyselected from the group consisting of C₁₋₆ aliphatic, halo, —OH,—N(R³)₂, cyano, —OR⁴, —C(O)R⁴, —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴,—N(R³)C(O)R⁴, —N(R³)CO₂R⁴, —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂,—N(R³)C(O)N(R³)₂, —OC(O)R⁴, —OC(O)N(R³)₂, phenyl, 5-6 memberedheterocyclyl and 5-6 membered heteroaryl, or two adjacent R⁵ on an aryl,cycloalkyl, or heteroaryl ring are taken together with their interveningatoms to form a 5-6 membered fused ring having 0-2 heteroatoms selectedfrom nitrogen, oxygen or sulfur; each R⁶ is independently selected fromthe group consisting of hydrogen, halo, C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄monoalkylamino and C₁₋₄ dialkylamino; and R⁷ is selected from the groupconsisting of hydrogen, halo, —OH, —N(R³)₂, cyano, —OR⁴, —C(O)R⁴,—CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴, —N(R³)C(O)R⁴, —N(R³)CO₂R⁴, —N(R³)SO₂R⁴,—C(O)N(R³)₂, —SO₂N(R³)₂, —N(R³)C(O)N(R³)₂, and —OC(O)R⁴.
 24. A methodfor inhibiting TGF-β activity in a subject, said method comprisingadministering to said subject a compound of claim
 1. 25. A methodcomprising inhibiting vascular smooth muscle cell proliferationassociated with procedural vascular trauma due to organ transplantation,vascular surgery, angioplasty, shunt placement, stent placement orvascular grafting, said method comprising administration to a mammal inneed thereof of a medical device coated with a biodegradable coatinghaving a compound of the formula:

or a pharmaceutically acceptable salt or a prodrug thereof, wherein Pgis hydrogen, alkyl or a nitrogen protecting group; n is an integer from0 to 3; each R¹ substituent is independently selected from the groupconsisting of —R², -T-R², and -V-T-R; each R² is independently selectedfrom the group consisting of C₁₋₃ aliphatic, hydroxy, —N(R³)₂, halo,cyano, —OR⁴, —C(O)R⁴, —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴, —N(R³)C(O)R⁴,—N(R³)CO₂R⁴, —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂, —N(R³)C(O)N(R³)₂,—OC(O)R⁴, phenyl which is optionally substituted with 1-3 R⁵, 5-6membered heterocyclyl which is optionally substituted with 1-3 R⁵, and5-6 membered heteroaryl which is optionally substituted with 1-3 R⁵;each T is independently C₁₋₅ alkylidene that is optionally interruptedby —O—, —C(O)—, —S—, —S(O)—, —S(O)₂—, or —N(R³)—; each V isindependently selected from the group consisting of —O—, —S—, —S(O)—,—S(O)₂—, —C(O)—, —N(R³)—, —N(R³)C(O)—, or —N(R³)C(O)₂—, —N(R³)S(O)₂—,—C(O)N(R³)—, —S(O)₂N(R³)—, —N(R³)C(O)N(R³)—, and —OC(O)—; each R³ isindependently selected from the group consisting of hydrogen, C₁₋₆aliphatic, —C(O)R⁴, —C(O)₂R⁴, and —SO₂R⁴, or two R³ on the same nitrogentogether with their intervening nitrogen form a 5-6 memberedheterocyclyl or heteroaryl ring having 1-3 ring heteroatoms selectedfrom nitrogen, oxygen or sulfur; each R⁴ is independently selected fromthe group consisting of C₁₋₆ aliphatic, phenyl or a 5-6 memberedheteroaryl or heterocyclyl having 1-3 ring heteroatoms selected fromnitrogen, oxygen or sulfur; R⁸ is selected from the group consisting of:(a) aryl, heteroaryl, and cycloalkyl, each of which is optionallysubstituted with 1-3 R⁵, and (b) alkyl; each R⁵ is independentlyselected from the group consisting of C₁₋₆ aliphatic, halo, —OH,—N(R³)₂, cyano, —OR⁴, —C(O)R⁴, —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴,—N(R³)C(O)R⁴, —N(R³)CO₂R⁴, —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂,—N(R³)C(O)N(R³)₂, —OC(O)R⁴, —OC(O)N(R³)₂, phenyl, 5-6 memberedheterocyclyl and 5-6 membered heteroaryl, or two adjacent R⁵ on an aryl,cycloalkyl, or heteroaryl ring are taken together with their interveningatoms to form a 5-6 membered fused ring having 0-2 heteroatoms selectedfrom nitrogen, oxygen or sulfur; each R⁶ is independently selected fromthe group consisting of hydrogen, halo, C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄monoalkylamino and C₁₋₄ dialkylamino; and R⁷ is selected from the groupconsisting of hydrogen, halo, —OH, —N(R³)₂, cyano, —OR⁴, —C(O)R⁴,—CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴, —N(R³)C(O)R⁴, —N(R³)CO₂R⁴, —N(R³)SO₂R⁴,—C(O)N(R³)₂, —SO₂N(R³)₂, —N(R³)C(O)N(R³)₂, and —OC(O)R⁴.
 26. A methodfor producing a 4-substituted quinazoline compound of the formula:

said method comprising contacting a 4-halogenated quinazoline compoundof the formula:

with an aminopyridinyl compound of the formula:

in the presence of a coupling catalyst under conditions sufficient toproduce said 4-substituted quinazoline compound, wherein n is an integerfrom 0 to 3; each R¹ substituent is independently selected from thegroup consisting of —R², -T-R², and -V-T-R; each R² is independentlyselected from the group consisting of C₁₋₃ aliphatic, hydroxy, —N(R³)₂,halo, cyano, —OR⁴, —C(O)R⁴, —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴,—N(R³)C(O)R⁴, —N(R³)CO₂R⁴, —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂,—N(R³)C(O)N(R³)₂, —OC(O)R⁴, phenyl which is optionally substituted with1-3 R⁵, 5-6 membered heterocyclyl which is optionally substituted with1-3 R⁵, and 5-6 membered heteroaryl which is optionally substituted with1-3 R⁵; each T is independently C₁₋₅ alkylidene that is optionallyinterrupted by —O—, —C(O)—, —S—, —S(O)—, —S(O)₂—, or —N(R³)—; each V isindependently selected from the group consisting of —O—, —S—, —S(O)—,—S(O)₂—, —C(O)—, —N(R³)—, —N(R³)C(O)—, or —N(R³)C(O)₂—, —N(R³)S(O)₂—,—C(O)N(R³)—, —S(O)₂N(R³)—, —N(R³)C(O)N(R³)—, and —OC(O)—; each R³ isindependently selected from the group consisting of hydrogen, C₁₋₆aliphatic, —C(O)R⁴, —C(O)₂R⁴, and —SO₂R⁴, or two R³ on the same nitrogentogether with their intervening nitrogen form a 5-6 memberedheterocyclyl or heteroaryl ring having 1-3 ring heteroatoms selectedfrom nitrogen, oxygen or sulfur; each R⁴ is independently selected fromthe group consisting of C₁₋₆ aliphatic, phenyl or a 5-6 memberedheteroaryl or heterocyclyl having 1-3 ring heteroatoms selected fromnitrogen, oxygen or sulfur; R⁸ is selected from the group consisting of:(a) aryl, heteroaryl, and cycloalkyl, each of which is optionallysubstituted with 1-3 R⁵, and (b) alkyl; each R⁵ is independentlyselected from the group consisting of C₁₋₆ aliphatic, halo, —OH,—N(R³)₂, cyano, —OR⁴, —C(O)R⁴, —CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴,—N(R³)C(O)R⁴, —N(R³)CO₂R⁴, —N(R³)SO₂R⁴, —C(O)N(R³)₂, —SO₂N(R³)₂,—N(R³)C(O)N(R³)₂, —OC(O)R⁴, —OC(O)N(R³)₂, phenyl, 5-6 memberedheterocyclyl and 5-6 membered heteroaryl, or two adjacent R⁵ on an aryl,cycloalkyl, or heteroaryl ring are taken together with their interveningatoms to form a 5-6 membered fused ring having 0-2 heteroatoms selectedfrom nitrogen, oxygen or sulfur; each R⁶ is independently selected fromthe group consisting of hydrogen, halo, C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄monoalkylamino and C₁₋₄ dialkylamino; and R⁷ is selected from the groupconsisting of hydrogen, halo, —OH, —N(R³)₂, cyano, —OR⁴, —C(O)R⁴,—CO₂R⁴, —SR⁴, —S(O)R⁴, —S(O)₂R⁴, —N(R³)C(O)R⁴, —N(R³)CO₂R⁴, —N(R³)SO₂R⁴,—C(O)N(R³)₂, —SO₂N(R³)₂, —N(R³)C(O)N(R³)₂, and —OC(O)R⁴.
 27. The methodof claim 26, wherein X is iodide.